3′3′-cyclic dinucleotides

ABSTRACT

The present invention relates to 3′3′-cyclic dinucleotides modified with a 3′-phosphonoalkyl bond and derivatives thereof, that can modulate the activity of the STING adaptor protein.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.62/654,058, filed Apr. 6, 2018, which is incorporated herein in theirentirety for all purposes.

SEQUENCE LISTING

This application contains a Sequence Listing which has been submittedelectronically in ASCII format and is hereby incorporated by referencein its entirety. Said ASCII copy, created on Jan. 23, 2019, is named052838_541001US_ST25.txt and is 6,914 bytes in size.

FIELD

The present invention relates to novel 3′3′-cyclic dinucleotides, e.g.,those modified with a 3′-phosphonoalkyl substituent and derivativesthereof. Such 3′3′-cyclic dinucleotides modulate the activity of theSTING adaptor protein (Stimulator of Interferon Genes).

Accordingly, the 3′3′-cyclic dinucleotides of the invention are believedto be useful in treating diseases in which modulation of STING adaptorprotein is beneficial, for example, inflammation, allergic andautoimmune diseases, cancer, viral infections caused by viruses such ashepatitis B and human immunodeficiency virus, and in immunogeniccompositions or vaccine adjuvants.

BACKGROUND

The innate immune system recognizes the presence of pathogen ordisruption of the homeostasis of the host by a battery of PatternRecognition Receptors (PRRs) which detect a small set of ligandsassociated with pathogens or damage. These ligands are generally calledPathogen Associated Molecular Patterns (PAMPs) or Damage AssociatedMolecular Patterns (DAMPs) (Takeuchi O et al, Cell, 2010:140, 805-820).A number of PRRs have been identified over past two decades includingToll-like receptors, retinoic acids inducible gene (RIG-I)-likereceptors, nucleotide-binding oligomerization domain-like (NOD)receptors, C-type lectin receptor and cytosolic DNA sensors (Brubaker SW et al, Annu Rev Immunol, 2015:33, 257-290). Recognition of PAMPs andDAMPs by PRRs ultimately leads to the upregulation of cytokines andchemokines, including interferons, and recruitment of immune cells tothe sites of infection. All of these processes slow down pathogenreplication and contribute to the development of adaptive immunity.

Cellular DNA is normally restricted to the nucleus and mitochondria ofhealthy cells. DNA present in cytosol, therefore, represents a signalindicating the presence of pathogen or disruption of the hosthomeostasis. The sensing of exogenous DNA is initiated by several DNAsensors such as DNA-dependent activator of IRFs (DAI) or DEAD boxpolypeptide 41 (DDX41). These DNA sensors signal via STING adaptorprotein (Stimulator Of Interferon Genes, also called STING, STINGprotein, TMEM173, MITA, or ERIS) (Unterholzner L, Immunology, 2013: 218,1312-1321) by recruiting protein kinase TBK1 that triggers activation ofthe transcription factors NF-κB (nuclear factor kappa B) and IRF-3(interferon regulatory factor 3). Activation of STING adaptor proteinultimately is believed to result in the release of type I and IIIinterferons as well as a variety of cytokines and chemokines such asinterleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) andinterferon-gamma (INF-γ).

Alternatively, STING protein can be activated by the second messengercyclic dinucleotides (CDNs) (Burdette et al. Nature 2011: 478, 515-518).CDNs with affinity to STING contain two purine nucleotide monophosphateslinked with either two 3′-5′ (3′3′-CDNs), two 2′-5′ (2′2′-CDNs) or 2′-5′and 3′-5′ phosphodiester bonds (2′3′-CDNs). The prototype 2′3′-cGAMP(c[G(2′,5′)pA(3′,5′)p]) is a product of the activation of host cGASprotein in the presence of pathogen or self dsDNA (Zhang et al,Molecular Cell 2013:51, 226-235).

The type I interferons (IFNs) are immune-regulatory cytokines that playa pivotal role in viral immunity. They can induce dendritic cell (DC)and macrophage maturation and activation (Galluci et al, Nat Med,1999:5, 1249-1255) and can promote T- and B-cell survival, activationand differentiation. Furthermore, the interferons are capable ofactivating numerous intracellular pathways that inhibit virusreplication. The clinical utility of type I interferons has beendemonstrated by their usefulness in treatment of chronic hepatitis B andC (Lin and Young, Cytokine Growth Factor Rev, 2014:25, 369-376).

In addition, interferons have shown utility in treatment of humancancers (Cohen et al, N Engl J Med, 2005:353, 2477-2490, Tsao et al, NEngl J Med, 2004:351, 998-1012). They can inhibit proliferation of tumorcells and may be synergistic with many approved anticancer agents.Furthermore, type-I-IFNs can act on immune cells to induce antitumorresponse (Musella et al, Oncoimmunology 2017:6:e1314424). Type I IFNsignaling was shown to be important in tumor-initiated T cell priming inmice. Animals lacking the IFN-α/13 receptor in dendritic cells wereunable to reject immunogenic tumors, and were defective in antigencross-presentation to CD8+ T cells (Fuertes et al, J Exp Med, 2011:208,2005-2016, Diamond et al, J Exp Med, 2011:208:1989-2003). Consistentwith these observations, intratumoral injection of STING agonists hasbeen recently shown to induce regression of established tumors in miceand to generate substantial systemic immune responses capable ofrejecting distant metastases and providing long-lived immunologic memory(Corrales et al, Cell Rep, 2015:11, 1018-1030).

CDNs are believed to promote priming of both cellular and humoralimmunity. For example, CDNs were shown to be an effective adjuvant inanimal models (Dubensky et al, Ther Adv Vaccines, 2013:1, 131-143.

Patent publications WO 2014/093936, WO 2014/189805, WO 2013/185052, US2014/03441976, WO 2015/077354, WO 2015/185565, WO 2016/145102, WO2017/093933, WO 2017/027646, WO 2017/027645, WO 2017/175156, WO2017/175147, WO 2017/123657, WO 2018/013908, WO 2018/013887, WO2018/009652, WO 2018/009648, and WO 2018/009466 disclose certain CDNsand their use in inducing an immune response.

It is an object of this invention to describe novel 3′3′-cyclicphosphonate dinucleotides and derivatives thereof that bind to andactivate the STING adaptor protein, thereby stimulating a signaltransduction pathway in order to induce interferons and othercytokines/chemokines. An advantage compared to the previously disclosedCDNs is believed to arise from the replacement of a phosphoester bondwith a phosphonoalkyl bond that is resistant toward hydrolysis byphosphodiesterases present in tissues and bodily fluids. Such compoundsmay find use as anti-viral or anti-cancer agents, as adjuvants invaccines, or as treatments of allergic or other inflammatory diseases.

BRIEF SUMMARY

In one embodiment, the present invention provides a structure accordingto Formula (I):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof,

wherein

-   -   L¹ is —O— or —K—C(R⁶R⁷)—;    -   L² is —K—C(R⁶R⁷)—;    -   Y¹ and Y² are each independently —O—, —S—, or —CH₂—;    -   X¹ and X³ are each independently OH, SH, OR¹³, SR¹³, or N(R¹³)₂;    -   X² and X⁴ are each independently O or S;    -   R¹, R⁵, R⁸ and R¹² are each independently H, CH₂F, CHF₂, CF₃,        CN, N₃, F, Cl, Br, I, COOR¹³, CON(R¹³)₂, CH₂OH, C₁-C₆ alkyl,        C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ substituted alkyl, C₂-C₆        substituted alkenyl, C₂-C₆ substituted alkynyl, OR¹³, SR¹³, or        N(R¹³)₂;    -   R², R³, R⁴, R⁹, R¹⁰ and R¹¹ are each independently H, OH, F, Cl,        Br, I, CN, N₃, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆        substituted alkyl, C₂-C₆ substituted alkenyl, C₂-C₆ substituted        alkynyl, OR¹³, SR¹³, or N(R¹³)₂;    -   R⁶ and R⁷ are each independently H, CH₂F, CHF₂, CF₃, CN, N₃, F,        Cl, Br, I, COOR¹³, CON(R¹³)₂, CH₂OH, CH₂N₃, OR¹³, SR¹³, N(R¹³)₂,        C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ substituted        alkyl, C₂-C₆ substituted alkenyl, C₂-C₆ substituted alkynyl,        C₃-C₇ cycloalkyl, C₂-C₁₀ heterocycloalkyl, C₂-C₁₀ substituted        heterocycloalkyl, C₆-C₁₀ aryl, C₆-C₁₀ substituted aryl, C₂-C₁₀        heteroaryl, or C₂-C₁₀ substituted heteroaryl;    -   each R¹³ is independently H, —C(═Z)R¹⁴, —C(═Z)OR¹⁴, —C(═Z)SR¹⁴,        —C(═Z)N(R¹⁴)₂, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆        substituted alkyl, C₂-C₆ substituted alkenyl, C₂-C₆ substituted        alkynyl, C₃-C₇ cycloalkyl, C₂-C₁₀ heterocycloalkyl, C₂-C₁₀        substituted heterocycloalkyl, C₆-C₁₀ aryl, C₆-C₁₀ substituted        aryl, C₂-C₁₀ heteroaryl, or C₂-C₁₀ substituted heteroaryl;    -   each R¹⁴ is independently H, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆        alkynyl, C₁-C₆ substituted alkyl, C₂-C₆ substituted alkenyl,        C₂-C₆ substituted alkynyl, C₃-C₇ cycloalkyl, C₂-C₁₀        heterocycloalkyl, C₂-C₁₀ substituted heterocycloalkyl, C₆-C₁₀        aryl, C₆-C₁₀ substituted aryl, C₂-C₁₀ heteroaryl, or C₂-C₁₀        substituted heteroaryl;    -   each Z is independently O, S, or NR¹³;    -   K is a variable that represents —O—, —S—, —S(O)—, —S(O)₂—, —NH—,        or —NR¹³—;    -   Base¹ and Base² are each independently:

wherein

-   -   A, A¹, A², A³ and A⁴ are each independently H, OH, SH, F, Cl,        Br, I, NH₂, OR¹³, SR¹³, NHR¹³, N(R¹³)₂, or R¹⁴; and    -   wherein the C₁-C₆ substituted alkyl, C₂-C₆ substituted alkenyl,        C₂-C₆ substituted alkynyl, C₂-C₁₀ substituted heterocycloalkyl,        C₆-C₁₀ substituted aryl, or C₂-C₁₀ substituted heteroaryl is        substituted by 1, 2, 3, or 4 substituents independently selected        from the group consisting of —OH, —SH, —NH₂, ═O, ═NH, ═S, ≡N,        halogen, —N₃, C₁₋₆ alkyl, C₁₋₆ alkoxy, —CN and —COOR_(p), where        R_(p) is hydrogen or C₁ to C₆ alkyl.

The present invention includes a pharmaceutical composition comprisingthe cyclic dinucleotide of Formula (I), or an enantiomer, hydrate,solvate or pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier, excipient, and/or diluent.

Also provided is a method of treating a disease or disorder, e.g., amethod of treating or preventing a viral infection, hepatitis B virusinfection, HIV infection, hyperproliferative disease or cancer,comprising administering to a human or animal in need thereof atherapeutically effective amount of a cyclic dinucleotide of Formula(I), or an enantiomer, hydrate, solvate or pharmaceutically acceptablesalt thereof, or a pharmaceutical composition of any of the foregoing.

Further provided is a method of enhancing the efficacy of a vaccine,comprising administering a therapeutically effective amount of a cyclicdinucleotide of Formula (I), or an enantiomer, hydrate, solvate orpharmaceutically acceptable salt thereof, or a pharmaceuticalcomposition of any of the foregoing.

Further provided is a method of modulating the activity of STING adaptorprotein to induce production of a type I interferon, cytokine and/orchemokine dependent on the STING adaptor protein, e.g., inducing a STINGadaptor protein-dependent type I interferon, cytokine or chemokine in ahuman or animal, comprising administering a therapeutically effectiveamount of a cyclic dinucleotide of Formula (I), or an enantiomer,hydrate, solvate or pharmaceutically acceptable salt thereof, or apharmaceutical composition of any of the foregoing.

DETAILED DESCRIPTION I. General

The invention provides novel cyclic dinucleotides, e.g., comprising atleast one phosphonoalkyl group, that bind to and modulate the activityof, e.g., activate, the STING protein.

II. Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art. A dash at the front or end of a chemical group is a matter ofconvenience to indicate the point of attachment to a parent moiety;chemical groups may be depicted with or without one or more dasheswithout losing their ordinary meaning. A prefix such as “C_(u-v)” or“C_(u)-C_(v)” indicates that the following group has from u to v carbonatoms, where u and v are integers. For example, “C₁₋₆ alkyl” or “C₁-C₆alkyl” indicates that the alkyl group has from 1 to 6 carbon atoms.

“Alkyl” is a linear or branched saturated monovalent hydrocarbon. Forexample, an alkyl group can have 1 to 10 carbon atoms (i.e., C₁₋₁₀alkyl) or 1 to 8 carbon atoms (i.e., C₁₋₈ alkyl) or 1 to 6 carbon atoms(i.e., C₁₋₆ alkyl) or 1 to 4 carbon atoms (i.e., C₁₋₄ alkyl). Examplesof alkyl groups include, but are not limited to, methyl (Me, —CH₃),ethyl (Et, —CH₂CH₃), 1-propyl (n-Pr, n-propyl, —CH₂CH₂CH₃), 2-propyl(i-Pr, i-propyl, —CH(CH₃)₂), 1-butyl (n-Bu, n-butyl, —CH₂CH₂CH₂CH₃),2-methyl-1-propyl (i-Bu, i-butyl, —CH₂CH(CH₃)₂), 2-butyl (s-Bu, s-butyl,—CH(CH₃)CH₂CH₃), 2-methyl-2-propyl (t-Bu, t-butyl, —C(CH₃)₃), 1-pentyl(n-pentyl, —CH₂CH₂CH₂CH₂CH₃), 2-pentyl (—CH(CH₃)CH₂CH₂CH₃), 3-pentyl(—CH(CH₂CH₃)₂), 2-methyl-2-butyl (—C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(—CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl (—CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(—CH₂CH(CH₃)CH₂CH₃), 1-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl(—CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl (—CH(CH₂CH₃)(CH₂CH₂CH₃)),2-methyl-2-pentyl (—C(CH₃)₂CH₂CH₂CH₃), 3-methyl-2-pentyl(—CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl (—CH(CH₃)CH₂CH(CH₃)₂),3-methyl-3-pentyl (—C(CH₃)(CH₂CH₃)₂), 2-methyl-3-pentyl(—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl (—C(CH₃)₂CH(CH₃)₂),3,3-dimethyl-2-butyl (—CH(CH₃)C(CH₃)₃, and octyl (—(CH₂)₇CH₃).

“Alkoxy” refers to the group —O-alkyl, where alkyl is as defined above.For example, C₁₋₄ alkoxy refers to an —O-alkyl group having 1 to 4carbons.

“Alkenyl” is a linear or branched monovalent hydrocarbon radical with atleast one carbon-carbon double bond. For example, an alkenyl group canhave 2 to 8 carbon atoms (i.e., C₂₋₈ alkenyl) or 2 to 6 carbon atoms(i.e., C₂₋₆ alkenyl) or 2 to 4 carbon atoms (i.e., C₂₋₄ alkenyl).Examples of alkenyl groups include, but are not limited to, ethenyl(—CH═CH₂), allyl (—CH₂CH═CH₂), and —CH₂—CH═CH—CH₃.

“Alkynyl” is a linear or branched monovalent hydrocarbon radical with atleast one carbon-carbon triple bond. For example, an alkynyl group canhave 2 to 8 carbon atoms (i.e., C₂₋₈ alkynyl) or 2 to 6 carbon atoms(i.e., C₂₋₆ alkynyl) or 2 to 4 carbon atoms (i.e., C₂₋₄ alkynyl).Examples of alkynyl groups include, but are not limited to, acetylenyl(—C≡CH), propargyl (—CH₂C≡CH), and —CH₂—C≡C—CH₃.

“Halo” or “halogen” as used herein refers to fluoro (—F), chloro (—Cl),bromo (—Br) and iodo (—I).

“Haloalkyl” as used herein refers to an alkyl as defined herein, whereinone or more hydrogen atoms of the alkyl are independently replaced by ahalo substituent, which may be the same or different. For example, C₁₋₄haloalkyl is a C₁₋₄ alkyl wherein one or more of the hydrogen atoms ofthe C₁₋₄ alkyl have been replaced by a halo substituent. Examples ofhaloalkyl groups include but are not limited to fluoromethyl,fluorochloromethyl, difluoromethyl, difluorochloromethyl,trifluoromethyl, 1,1,1-trifluoroethyl and pentafluoroethyl.

“Aryl” as used herein refers to a single all carbon aromatic ring or amultiple condensed all carbon ring system wherein at least one of therings is aromatic. For example, in certain embodiments, an aryl grouphas 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 12 carbon atoms.Aryl includes a phenyl radical. Aryl also includes multiple condensedring systems (e.g., ring systems comprising 2, 3 or 4 rings) havingabout 9 to 20 carbon atoms in which at least one ring is aromatic andwherein the other rings may be aromatic or not aromatic (i.e.,carbocycle). Such multiple condensed ring systems are optionallysubstituted with one or more (e.g., 1, 2 or 3) oxo groups on anycarbocycle portion of the multiple condensed ring system. The rings ofthe multiple condensed ring system can be connected to each other viafused, spiro and bridged bonds when allowed by valency requirements. Itis also to be understood that when reference is made to a certainatom-range membered aryl (e.g., 6-10 membered aryl), the atom range isfor the total ring atoms of the aryl. For example, a 6-membered arylwould include phenyl and a 10-membered aryl would include naphthyl and1,2,3,4-tetrahydronaphthyl. Non-limiting examples of aryl groupsinclude, but are not limited to, phenyl, indenyl, naphthyl,1,2,3,4-tetrahydronaphthyl, anthracenyl, and the like.

“Heteroaryl” as used herein refers to a single aromatic ring that has atleast one atom other than carbon in the ring, wherein the atom isselected from the group consisting of oxygen, nitrogen and sulfur;“heteroaryl” also includes multiple condensed ring systems that have atleast one such aromatic ring, which multiple condensed ring systems arefurther described below. Thus, “heteroaryl” includes single aromaticrings of from about 1 to 6 carbon atoms and about 1-4 heteroatomsselected from the group consisting of oxygen, nitrogen and sulfur. Thesulfur and nitrogen atoms may also be present in an oxidized formprovided the ring is aromatic. Exemplary heteroaryl ring systems includebut are not limited to pyridyl, pyrimidinyl, oxazolyl or furyl.“Heteroaryl” also includes multiple condensed ring systems (e.g., ringsystems comprising 2, 3 or 4 rings) wherein a heteroaryl group, asdefined above, is condensed with one or more rings selected fromheteroaryls (to form for example 1,8-naphthyridinyl), heterocycles, (toform for example 1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocycles (toform for example 5,6,7,8-tetrahydroquinolyl) and aryls (to form forexample indazolyl) to form the multiple condensed ring system. Thus, aheteroaryl (a single aromatic ring or multiple condensed ring system)has about 1-20 carbon atoms and about 1-6 heteroatoms within theheteroaryl ring. Such multiple condensed ring systems may be optionallysubstituted with one or more (e.g., 1, 2, 3 or 4) oxo groups on thecarbocycle or heterocycle portions of the condensed ring. The rings ofthe multiple condensed ring system can be connected to each other viafused, spiro and bridged bonds when allowed by valency requirements. Itis to be understood that the individual rings of the multiple condensedring system may be connected in any order relative to one another. It isto be understood that the point of attachment for a heteroaryl orheteroaryl multiple condensed ring system can be at any suitable atom ofthe heteroaryl or heteroaryl multiple condensed ring system including acarbon atom and a heteroatom (e.g., a nitrogen). It also to beunderstood that when a reference is made to a certain atom-rangemembered heteroaryl (e.g., a 5 to 10 membered heteroaryl), the atomrange is for the total ring atoms of the heteroaryl and includes carbonatoms and heteroatoms. For example, a 5-membered heteroaryl wouldinclude a thiazolyl and a 10-membered heteroaryl would include aquinolinyl. Exemplary heteroaryls include but are not limited topyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl,thienyl, indolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, furyl,oxadiazolyl, thiadiazolyl, quinolyl, isoquinolyl, benzothiazolyl,benzoxazolyl, indazolyl, quinoxalyl, quinazolyl,5,6,7,8-tetrahydroisoquinolinyl benzofuranyl, benzimidazolyl,thianaphthenyl, pyrrolo[2,3-b]pyridinyl, quinazolinyl-4(3H)-one, andtriazolyl.

“Cycloalkyl” refers to a single saturated or partially unsaturated allcarbon ring having 3 to 20 annular carbon atoms (i.e., C₃₋₂₀cycloalkyl), for example from 3 to 12 annular atoms, for example from 3to 10 annular atoms, or 3 to 8 annular atoms, or 3 to 6 annular atoms,or 3 to 5 annular atoms, or 3 to 4 annular atoms. The term “cycloalkyl”also includes multiple condensed, saturated and partially unsaturatedall carbon ring systems (e.g., ring systems comprising 2, 3 or 4carbocyclic rings). Accordingly, cycloalkyl includes multicycliccarbocyles such as a bicyclic carbocycles (e.g., bicyclic carbocycleshaving about 6 to 12 annular carbon atoms such as bicyclo[3.1.0]hexaneand bicyclo[2.1.1]hexane), and polycyclic carbocycles (e.g tricyclic andtetracyclic carbocycles with up to about 20 annular carbon atoms). Therings of a multiple condensed ring system can be connected to each othervia fused, spiro and bridged bonds when allowed by valency requirements.Non-limiting examples of monocyclic cycloalkyl include cyclopropyl,cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl,1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl and1-cyclohex-3-enyl.

“Heterocyclyl” or “heterocycle” or “heterocycloalkyl” as used hereinrefers to a single saturated or partially unsaturated non-aromatic ringor a non-aromatic multiple ring system that has at least one heteroatomin the ring (i.e., at least one annular heteroatom selected from oxygen,nitrogen, and sulfur). Unless otherwise specified, a heterocyclyl grouphas from 3 to about 20 annular atoms, for example from 3 to 12 annularatoms, for example from 3 to 10 annular atoms, or 3 to 8 annular atoms,or 3 to 6 annular atoms, or 3 to 5 annular atoms, or 4 to 6 annularatoms, or 4 to 5 annular atoms. Thus, the term includes single saturatedor partially unsaturated rings (e.g., 3, 4, 5, 6 or 7-membered rings)having from about 1 to 6 annular carbon atoms and from about 1 to 3annular heteroatoms selected from the group consisting of oxygen,nitrogen and sulfur in the ring. The rings of the multiple condensedring (e.g. bicyclic heterocyclyl) system can be connected to each othervia fused, spiro and bridged bonds when allowed by valency requirements.Heterocycles include, but are not limited to, azetidine, aziridine,imidazolidine, morpholine, oxirane (epoxide), oxetane, thietane,piperazine, piperidine, pyrazolidine, piperidine, pyrrolidine,pyrrolidinone, tetrahydrofuran, tetrahydrothiophene, dihydropyridine,tetrahydropyridine, quinuclidine, 2-oxa-6-azaspiro[3.3]heptan-6-yl,6-oxa-1-azaspiro[3.3]heptan-1-yl, 2-thia-6-azaspiro[3.3]heptan-6-yl,2,6-diazaspiro[3.3]heptan-2-yl, 2-azabicyclo[3.1.0]hexan-2-yl,3-azabicyclo[3.1.0]hexanyl, 2-azabicyclo[2.1.1]hexanyl,2-azabicyclo[2.2.1]heptan-2-yl, 4-azaspiro[2.4]heptanyl,5-azaspiro[2.4]heptanyl, and the like.

“Oxo” as used herein refers to ═O.

“Substituted” as used herein refers to substitution with one or moresubstituents (e.g., 1, 2, 3, or 4 or more) selected from the groupconsisting of —OH, —SH, —NH₂, ═O, ═NH, ═S, ≡N, halogen, —N₃, C₁₋₆ alkyl,C₁₋₆ alkoxy, —CN and —COOR_(p), where R_(p) is hydrogen or C₁ to C₆alkyl.

A “compound of the present disclosure” includes compounds disclosedherein, for example a compound of the present disclosure includescompounds of Formula (I), (I-1), (I-2), (Ia-1), (Ia-2), (Ib), (Ic),(Id), (Ie), (If), and/or (Ig), including the compounds of the Examples.

“Treatment” or “treat” or “treating” as used herein refers to anapproach for obtaining beneficial or desired results. For purposes ofthe present disclosure, beneficial or desired results include, but arenot limited to, alleviation of a symptom and/or diminishment of theextent of a symptom and/or preventing a worsening of a symptomassociated with a disease or condition. In one embodiment, “treatment”or “treating” includes one or more of the following: a) inhibiting thedisease or condition (e.g., decreasing one or more symptoms resultingfrom the disease or condition, and/or diminishing the extent of thedisease or condition); b) slowing or arresting the development of one ormore symptoms associated with the disease or condition (e.g.,stabilizing the disease or condition, delaying the worsening orprogression of the disease or condition); and c) relieving the diseaseor condition, e.g., causing the regression of clinical symptoms,ameliorating the disease state, delaying the progression of the disease,increasing the quality of life, and/or prolonging survival.

“Delaying” as used herein refers to development of a disease orcondition means to defer, hinder, slow, retard, stabilize and/orpostpone development of the disease or condition. This delay can be ofvarying lengths of time, depending on the history of the disease and/orindividual being treated. As is evident to one skilled in the art, asufficient or significant delay can, in effect, encompass prevention, inthat the individual does not develop the disease or condition.

“Prevent” or “prevention” or “preventing” as used herein refers to aregimen that protects against the onset of the disease or disorder suchthat the clinical symptoms of the disease do not develop. Thus,“prevention” relates to administration of a therapy (e.g.,administration of a therapeutic substance) to a subject before signs ofthe disease are detectable in the subject (e.g., administration of atherapeutic substance to a subject in the absence of detectableinfectious agent (e.g., virus) in the subject). The subject may be anindividual at risk of developing the disease or disorder, such as anindividual who has one or more risk factors known to be associated withdevelopment or onset of the disease or disorder. Thus, in certainembodiments, the term “preventing HBV infection” refers to administeringto a subject who does not have a detectable HBV infection an anti-HBVtherapeutic substance. It is understood that the subject for anti-HBVpreventative therapy may be an individual at risk of contracting the HBVvirus. It is also understood that prevention does not require a 100%success rate. In some instances, prevention may be understood as areduction of the risk of infection, but not a complete elimination theoccurrence of an infection.

“Modulation” or “modulating” the activity of a protein, e.g., a STINGadaptor protein, as used herein refers to alteration of the activitysuch that the activity increases or decreases. In some embodiments, themodulation increases the activity.

As used herein, the term “viral infection” describes a diseased state inwhich a virus invades healthy cells, uses the cell's reproductivemachinery to multiply or replicate and ultimately lyse the cellresulting in cell death, release of viral particles and the infection ofother cells by the newly produced progeny viruses. Latent infection bycertain viruses is also a possible result of viral infection.

As used herein, the term “enhancing” refers to any form of increase inthe immunogenic activity of an effective dosage of a vaccine as a resultof administering to an animal or a human a therapeutically effectivedose of a compound of the present disclosure, e.g., a cyclicdinucleotide of Formula (I), (I-1), (I-2), (Ia-1), (Ia-2), (Ib), (Ic),(Id), (Ie), (If), and/or (Ig), wherein said compound is administered atany time prior to, simultaneous with, or just after administration tothe same animal or human of the effective dosage of a vaccine.

“Animal” as used herein refers to a mammal, for example, a domesticanimal such as a pig, a cow, a horse, a dog, a cat, a rat, or a mouse,or a non-human primate such as a cynomolgus monkey or chimpanzee.

“At risk individual” as used herein refers to an individual who is atrisk of developing a condition to be treated. An individual “at risk”may or may not have detectable disease or condition, and may or may nothave displayed detectable disease prior to the treatment of methodsdescribed herein. “At risk” denotes that an individual has one or moreso-called risk factors, which are measurable parameters that correlatewith development of a disease or condition and are known in the art. Anindividual having one or more of these risk factors has a higherprobability of developing the disease or condition than an individualwithout these risk factor(s).

“Therapeutically effective amount” or “effective amount” as used hereinrefers to an amount that is effective to elicit the desired biologicalor medical response, including the amount of a compound that, whenadministered to a subject for treating a disease, is sufficient toeffect such treatment for the disease. The effective amount will varydepending on the compound, the disease, and its severity and the age,weight, etc., of the subject to be treated. The effective amount caninclude a range of amounts. As is understood in the art, an effectiveamount may be in one or more doses, i.e., a single dose or multipledoses may be required to achieve the desired treatment endpoint. Aneffective amount may be considered in the context of administering oneor more therapeutic agents, and a single agent may be considered to begiven in an effective amount if, in conjunction with one or more otheragents, a desirable or beneficial result may be or is achieved. Suitabledoses of any co-administered compounds may optionally be lowered due tothe combined action (e.g., additive or synergistic effects) of thecompounds.

“Pharmaceutically acceptable excipient” includes without limitation anyadjuvant, carrier, excipient, glidant, sweetening agent, diluent,preservative, dye/colorant, flavor enhancer, surfactant, wetting agent,dispersing agent, suspending agent, stabilizer, isotonic agent, solvent,or emulsifier which has been approved by the United States Food and DrugAdministration as being acceptable for use in humans or domesticanimals.

“Co-administration” as used herein refers to administration of unitdosages of the compounds disclosed herein before or after administrationof unit dosages of one or more additional therapeutic agents, forexample, administration of the compound disclosed herein within seconds,minutes, or hours of the administration of one or more additionaltherapeutic agents. For example, in some embodiments, a unit dose of acompound of the present disclosure is administered first, followedwithin seconds or minutes by administration of a unit dose of one ormore additional therapeutic agents. Alternatively, in other embodiments,a unit dose of one or more additional therapeutic agents is administeredfirst, followed by administration of a unit dose of a compound of thepresent disclosure within seconds or minutes. In some embodiments, aunit dose of a compound of the present disclosure is administered first,followed, after a period of hours (e.g., 1-12 hours), by administrationof a unit dose of one or more additional therapeutic agents. In otherembodiments, a unit dose of one or more additional therapeutic agents isadministered first, followed, after a period of hours (e.g., 1-12hours), by administration of a unit dose of a compound of the presentdisclosure. Co-administration of a compound disclosed herein with one ormore additional therapeutic agents generally refers to simultaneous orsequential administration of a compound disclosed herein and one or moreadditional therapeutic agents, such that therapeutically effectiveamounts of each agent are present in the body of the patient.

Provided are also pharmaceutically acceptable salts, hydrates, solvates,tautomeric forms, polymorphs, and prodrugs of the compounds describedherein. “Pharmaceutically acceptable” or “physiologically acceptable”refer to compounds, salts, compositions, dosage forms and othermaterials which are useful in preparing a pharmaceutical compositionthat is suitable for veterinary or human pharmaceutical use.

The compounds of described herein may be prepared and/or formulated aspharmaceutically acceptable salts or when appropriate as a free base.Pharmaceutically acceptable salts are non-toxic salts of a free baseform of a compound that possesses the desired pharmacological activityof the free base. These salts may be derived from inorganic or organicacids or bases. For example, a compound that contains a basic nitrogenmay be prepared as a pharmaceutically acceptable salt by contacting thecompound with an inorganic or organic acid. Non-limiting examples ofpharmaceutically acceptable salts include sulfates, pyrosulfates,bisulfates, sulfites, bisulfites, phosphates, monohydrogen-phosphates,dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides,bromides, iodides, acetates, propionates, decanoates, caprylates,acrylates, formates, isobutyrates, caproates, heptanoates, propiolates,oxalates, malonates, succinates, suberates, sebacates, fumarates,maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates,chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates,methoxybenzoates, phthalates, sulfonates, methylsulfonates,propylsulfonates, besylates, xylenesulfonates, naphthalene-1-sulfonates,naphthalene-2-sulfonates, phenylacetates, phenylpropionates,phenylbutyrates, citrates, lactates, γ-hydroxybutyrates, glycolates,tartrates, and mandelates. Lists of other suitable pharmaceuticallyacceptable salts are found in Remington: The Science and Practice ofPharmacy, 21^(st) Edition, Lippincott Wiliams and Wilkins, Philadelphia,Pa., 2006.

Examples of “pharmaceutically acceptable salts” of the compoundsdisclosed herein also include salts derived from an appropriate base,such as an alkali metal (for example, sodium, potassium), an alkalineearth metal (for example, magnesium), ammonium and NX₄ ⁺ (wherein X isC₁-C₄ alkyl). Also included are base addition salts, such as sodium orpotassium salts.

Provided are also compounds described herein or pharmaceuticallyacceptable salts, isomers, or a mixture thereof, in which from 1 to nhydrogen atoms attached to a carbon atom may be replaced by a deuteriumatom or D, in which n is the number of hydrogen atoms in the molecule.As known in the art, the deuterium atom is a non-radioactive isotope ofthe hydrogen atom. Such compounds may increase resistance to metabolism,and thus may be useful for increasing the half-life of the compoundsdescribed herein or pharmaceutically acceptable salts, isomer, or amixture thereof when administered to a mammal. See, e.g., Foster,“Deuterium Isotope Effects in Studies of Drug Metabolism”, TrendsPharmacol. Sci., 5(12):524-527 (1984). Such compounds are synthesized bymeans well known in the art, for example by employing starting materialsin which one or more hydrogen atoms have been replaced by deuterium.

Examples of isotopes that can be incorporated into the disclosedcompounds also include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, fluorine, chlorine, and iodine, such as ²H, ³H, ¹¹C, ¹³C,¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³¹P, ³²P, 35S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I,respectively. Substitution with positron emitting isotopes, such as ¹¹C,¹⁸F, ¹⁵O and ¹³N, can be useful in Positron Emission Topography (PET)studies for examining substrate receptor occupancy. Isotopically-labeledcompounds of Formula (I) can generally be prepared by conventionaltechniques known to those skilled in the art or by processes analogousto those described in the Examples as set out below using an appropriateisotopically-labeled reagent in place of the non-labeled reagentpreviously employed.

The compounds of the embodiments disclosed herein, or theirpharmaceutically acceptable salts may contain one or more asymmetriccenters and may thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids.The present disclosure is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optically active (+) and(−), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiralsynthons or chiral reagents, or resolved using conventional techniques,for example, chromatography and fractional crystallization. Conventionaltechniques for the preparation/isolation of individual enantiomersinclude chiral synthesis from a suitable optically pure precursor orresolution of the racemate (or the racemate of a salt or derivative)using, for example, chiral high pressure liquid chromatography (HPLC).When the compounds described herein contain olefinic double bonds orother centres of geometric asymmetry, and unless specified otherwise, itis intended that the compounds include both E and Z geometric isomers.Likewise, all tautomeric forms are also intended to be included. Wherecompounds are represented in their chiral form, it is understood thatthe embodiment encompasses, but is not limited to, the specificdiastereomerically or enantiomerically enriched form. Where chirality isnot specified but is present, it is understood that the embodiment isdirected to either the specific diastereomerically or enantiomericallyenriched form; or a racemic or scalemic mixture of such compound(s). Asused herein, “scalemic mixture” is a mixture of stereoisomers at a ratioother than 1:1.

“Stereoisomer” as used herein refers to a compound made up of the sameatoms bonded by the same bonds but having different three-dimensionalstructures, which are not interchangeable. The present disclosurecontemplates various stereoisomers and mixtures thereof and includes“enantiomers”, which refers to two stereoisomers whose molecules arenon-superimposable mirror images of one another.

“Tautomer” as used herein refers to a proton shift from one atom of amolecule to another atom of the same molecule. The present disclosureincludes tautomers of any said compounds.

“Solvate” as used herein refers to the result of the interaction of asolvent and a compound. Solvates of salts of the compounds describedherein are also provided. Hydrates of the compounds described herein arealso provided.

“Hydrate” as used herein refers to a compound of the disclosure that ischemically associated with one or more molecules of water.

“Prodrug” as used herein refers to a derivative of a drug that uponadministration to the human body is converted to the parent drugaccording to some chemical or enzymatic pathway.

III. Compounds

The invention includes cyclic dinucleotides of Formula (I),

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof, wherein

-   -   L¹ is —O— or —K—C(R⁶R⁷)—;    -   L² is —K—C(R⁶R⁷)—;    -   Y¹ and Y² are each independently —O—, —S—, or —CH₂—;    -   X¹ and X³ are each independently OH, SH, OR¹³, SR¹³, or N(R¹³)₂;    -   X² and X⁴ are each independently O or S;    -   R¹, R⁵, R⁸ and R¹² are each independently H, CH₂F, CHF₂, CF₃,        CN, N₃, F, Cl, Br, I, COOR¹³, CON(R¹³)₂, CH₂OH, C₁-C₆ alkyl,        C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ substituted alkyl, C₂-C₆        substituted alkenyl, C₂-C₆ substituted alkynyl, OR¹³, SR¹³, or        N(R¹³)₂;    -   R², R³, R⁴, R⁹, R¹⁰ and R¹¹ are each independently H, OH, F, Cl,        Br, I, CN, N₃, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆        substituted alkyl, C₂-C₆ substituted alkenyl, C₂-C₆ substituted        alkynyl, OR¹³, SR¹³, or N(R¹³)₂;    -   R⁶ and R⁷ are each independently H, CH₂F, CHF₂, CF₃, CN, N₃, F,        Cl, Br, I, COOR¹³, CON(R¹³)₂, CH₂OH, CH₂N₃, OR¹³, SR¹³, N(R¹³)₂,        C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ substituted        alkyl, C₂-C₆ substituted alkenyl, C₂-C₆ substituted alkynyl,        C₃-C₇ cycloalkyl, C₂-C₁₀ heterocycloalkyl, C₂-C₁₀ substituted        heterocycloalkyl, C₆-C₁₀ aryl, C₆-C₁₀ substituted aryl, C₂-C₁₀        heteroaryl, or C₂-C₁₀ substituted heteroaryl;    -   each R¹³ is independently H, —C(═Z)R¹⁴, —C(═Z)OR¹⁴, —C(═Z)SR¹⁴,        —C(═Z)N(R¹⁴)₂, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆        substituted alkyl, C₂-C₆ substituted alkenyl, C₂-C₆ substituted        alkynyl, C₃-C₇ cycloalkyl, C₂-C₁₀ heterocycloalkyl, C₂-C₁₀        substituted heterocycloalkyl, C₆-C₁₀ aryl, C₆-C₁₀ substituted        aryl, C₂-C₁₀ heteroaryl, or C₂-C₁₀ substituted heteroaryl;    -   each R¹⁴ is independently H, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆        alkynyl, C₁-C₆ substituted alkyl, C₂-C₆ substituted alkenyl,        C₂-C₆ substituted alkynyl, C₃-C₇ cycloalkyl, C₂-C₁₀        heterocycloalkyl, C₂-C₁₀ substituted heterocycloalkyl, C₆-C₁₀        aryl, C₆-C₁₀ substituted aryl, C₂-C₁₀ heteroaryl, or C₂-C₁₀        substituted heteroaryl;    -   each Z is independently O, S, or NR¹³;    -   K is a variable that represents —O—, —S—, —S(O)—, —S(O)₂—, —NH—,        or —NR¹³—;    -   Base¹ and Base² are each independently:

wherein

-   -   A, A¹, A², A³ and A⁴ are each independently H, OH, SH, F, Cl,        Br, I, NH₂, OR¹³, SR¹³, NHR¹³, N(R¹³)₂, or R¹⁴; and    -   wherein the C₁-C₆ substituted alkyl, C₂-C₆ substituted alkenyl,        C₂-C₆ substituted alkynyl, C₂-C₁₀ substituted heterocycloalkyl,        C₆-C₁₀ substituted aryl, or C₂-C₁₀ substituted heteroaryl is        substituted by 1, 2, 3, or 4 substituents independently selected        from the group consisting of —OH, —SH, —NH₂, ═O, ═NH, ═S, ≡N,        halogen, —N₃, C₁₋₆ alkyl, C₁₋₆ alkoxy, —CN and —COOR_(p), where        R_(p) is hydrogen or C₁ to C₆ alkyl.

In some embodiments, the compound is a compound of Formula (I):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof,

wherein

-   -   L¹ is —O— or —K—C(R⁶R⁷)—;    -   L² is —K—C(R⁶R⁷)—;    -   Y¹ and Y² are each independently —O—, —S—, or —CH₂—;    -   X¹ and X³ are each independently OH, SH, OR¹³, SR¹³, or N(R¹³)₂;    -   X² and X⁴ are each independently O or S;    -   R¹, R⁵, R⁸ and R¹² are each independently H, CH₂F, CHF₂, CF₃,        CN, N₃, F, Cl, Br, I, COOR¹³, CON(R¹³)₂, CH₂OH, C₁-C₆ alkyl,        C₂-C₆ alkenyl, C₂-C₆ alkynyl, OR¹³, SR¹³, or N(R¹³)₂;    -   R², R³, R⁴, R⁹, R¹⁰ and R¹¹ are each independently H, OH, F, Cl,        Br, I, CN, N₃, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, OR¹³,        SR¹³, or N(R¹³)₂;    -   R⁶ and R⁷ are each independently H, CH₂F, CHF₂, CF₃, CN, N₃, F,        Cl, Br, I, COOR¹³, CON(R¹³)₂, CH₂OH, CH₂N₃, OR¹³, SR¹³, N(R¹³)₂,        C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl,        C₂-C₁₀ heterocycloalkyl, C₆-C₁₀ aryl, or C₂-C₁₀ heteroaryl;    -   each R¹³ is independently H, —C(═Z)R¹⁴, —C(═Z)OR¹⁴, —C(═Z)SR¹⁴,        —C(═Z)N(R¹⁴)₂, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₇        cycloalkyl, C₂-C₁₀ heterocycloalkyl, C₆-C₁₀ aryl, or C₂-C₁₀        heteroaryl;    -   each R¹⁴ is independently H, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆        alkynyl, C₃-C₇ cycloalkyl, C₂-C₁₀ heterocycloalkyl, C₆-C₁₀ aryl,        or C₂-C₁₀ heteroaryl;    -   each Z is independently O, S, or NR¹³;    -   K is a variable that represents —O—, —S—, —S(O)—, —S(O)₂—, —NH—,        or —NR³—;    -   Base¹ and Base² are each independently:

wherein

-   -   A, A¹, A², A³ and A⁴ are each independently H, OH, SH, F, Cl,        Br, I, NH₂, OR¹³, SR¹³, NHR¹³, N(R¹³)₂, or R¹⁴.

In some embodiments, Y¹ and Y² are O.

In some embodiments, X² and X⁴ are O, and X¹ and X³ are eachindependently OH or OR¹³. In some embodiments, X² and X⁴ are O, and X¹and X³ are OH.

In some embodiments, L¹ is —O—. In some embodiments, L¹ is —O—C(R⁶R⁷)—.In some embodiments, L¹ is —O—CH₂—.

In some embodiments, L² is —O—C(R⁶R⁷)—. In some embodiments, L² is—O—CH₂—.

In some embodiments, L¹ is —O— or —O—C(R⁶R⁷)—, and L² is —O—C(R⁶R⁷)—. Insome embodiments, L¹ is —O— or —O—CH₂—, and L² is —O—CH₂—. In someembodiments, L¹ is —O—, and L² is —O—CH₂—. In some embodiments, L¹ is—O—CH₂—, and L² is —O—CH₂—.

In some embodiments, R¹, R⁵, R⁸ and R¹² are each independently H, CH₂F,CHF₂, CF₃, CN, N₃, F, Cl, Br, I, COOR¹³, CON(R¹³)₂, CH₂OH, C₁₋₆ alkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, OR¹³, SR¹³, or N(R¹³)₂. In some embodiments,R¹, R⁵, R⁸ and R¹² are each independently H, CN, N₃, F, Cl, Br, I,COOR¹³, CON(R¹³)₂, CH₂OH, C₁₋₃ haloalkyl, C₁₋₆ alkyl, OR¹³, SR¹³, orN(R¹³)₂. In some embodiments, R¹, R⁵, R⁸ and R¹² are each H.

In some embodiments, R², R³, R⁴, R⁹, R¹⁰ and R¹¹ are each independentlyH, OH, F, Cl, Br, I, CN, N₃, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,OR¹³, SR¹³, or N(R¹³)₂. In some embodiments, R², R³, R⁴, R⁹, R¹⁰ and R¹¹are each independently H, OH, F, Cl, Br, I, CN, N₃, C₁-C₆ alkyl, OR¹³,SR¹³, or N(R¹³)₂.

In some embodiments, R², R⁴, R⁹, and R¹¹ are each independently H, OH,F, Cl, Br, I, CN, or N₃. In some embodiments, R², R⁴, R⁹, and R¹¹ areeach H.

In some embodiments, R⁶ and R⁷ are each independently H, CH₂F, CHF₂,CF₃, CN, N₃, F, Cl, Br, I, COOR¹³, CON(R¹³)₂, CH₂OH, CH₂N₃, OR¹³, SR¹³,N(R¹³)₂, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl,C₂-C₁₀ heterocycloalkyl, C₆-C₁₀ aryl, or C₂-C₁₀ heteroaryl. In someembodiments, R⁶ and R⁷ are each independently H, CN, N₃, F, Cl, Br, I,COOR¹³, CON(R¹³)₂, CH₂OH, CH₂N₃, OR¹³, SR¹³, N(R¹³)₂, C₁-C₃ haloalkyl,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl, C₂-C₁₀heterocycloalkyl, C₆-C₁₀ aryl, or C₂-C₁₀ heteroaryl. In someembodiments, R⁶ and R⁷ are each independently H, C₁-C₃ haloalkyl, CN,N₃, F, Cl, Br, and I. In some embodiments, R⁶ and R⁷ are eachindependently H, CH₂F, CHF₂, CF₃, CN, N₃, F, Cl, Br, and I. In someembodiments, R⁶ and R⁷ are each H.

In some embodiments, each R¹³ is independently H, —C(═Z)R¹⁴, —C(═Z)OR¹⁴,—C(═Z)SR¹⁴, —C(═Z)N(R¹⁴)₂, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₃-C₇ cycloalkyl, C₂-C₁₀ heterocycloalkyl, C₆-C₁₀ aryl, or C₂-C₁₀heteroaryl. In some embodiments, each R¹³ is independently H or C₁-C₆alkyl.

In some embodiments, each R¹⁴ is independently H, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₃-C₇ cycloalkyl, C₂-C₁₀ heterocycloalkyl,C₆-C₁₀ aryl, or C₂-C₁₀ heteroaryl. In some embodiments, each R¹⁴ isindependently H or C₁-C₆ alkyl.

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (I-1):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof,

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (I-2):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof,

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (Ia-1):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments of the structure of Formula (I) and/or (Ia-1), L¹ is—O— or —O—C(R⁶R⁷)—, and L² is —O—C(R⁶R⁷)—. In some embodiments, L¹ is—O— or —O—CH₂—, and L² is —O—CH₂—. In some embodiments, L¹ is —O—, andL² is —O—CH₂—. In some embodiments, L¹ is —O—CH₂—, and L² is —O—CH₂—.

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (Ia-2):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments of the structure of Formula (Ia-2), L¹ is —O— or—O—C(R⁶R⁷)—.

In some embodiments, L¹ is —O— or —O—CH₂—. In some embodiments, L¹ is—O—. In some embodiments, L¹ is —O—CH₂—.

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (Ib):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (Ic):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (Id):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (Ie):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (If):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula (I) has astructure of Formula (Ig):

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments of the structure of Formula (I), (I-1), (I-2),(Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig), Base¹ andBase² are each independently:

In some embodiments, Base¹ and Base² are each independently:

In some embodiments, Base¹ and Base² are each independently:

In some embodiments, Base¹ and Base² are each independently:

In some embodiments, Base¹ and Base² are each independently:

In some embodiments, Base¹ is

and Base² is

In some embodiments, Base¹ is

and Base² is

In some embodiments, Base¹ and Base² are each

In some embodiments, Base¹ and Base² are each

In some embodiments of the structure of Formula ((I), (I-1), (I-2),(Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig), A, A¹, A², A³and A⁴ are each independently H, OH, SH, F, Cl, Br, I, NH₂.

In some embodiments of the structure of Formula (I), (I-1), (I-2),(Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig), A¹, A², andA³ are each independently H, OH, SH, F, Cl, Br, I, NH₂. In someembodiments, A¹, A², and A³ are each independently H, OH, and NH₂.

In some embodiments of the structure of Formula (I), (I-1), (I-2),(Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig), R³ and R¹⁰are each independently H, OH, F, Cl, Br, I, CN, N₃, or OR¹³. In someembodiments, R³ and R¹⁰ are each independently H, OH, F, Cl, Br, orOR¹³. In some such embodiments, R¹³ is C₁-C₆ alkyl. In some embodiments,R³ and R¹⁰ are each independently H, OH or F. In some embodiments, R³and R¹⁰ are each independently OH or F. In some embodiments, R³ is OH,and R¹⁰ is F. In some embodiments, R³ is F, and R¹⁰ is OH. In someembodiments, R³ is H, and R¹⁰ is F. In some embodiments, R³ is F, andR¹⁰ is H. In some embodiments, R³ and R¹⁰ are each OH. In someembodiments, R³ and R¹⁰ are each F.

In some embodiments, the cyclic dinucleotide of Formula (I), (I-1),(I-2), (Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig), hasthe structure:

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula (I), (I-1),(I-2), (Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig) has thestructure:

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula (I), (I-1),(I-2), (Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig) has thestructure:

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula (I), (I-1),(I-2), (Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig) has thestructure:

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula ((I), (I-1),(I-2), (Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig) has thestructure:

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

In some embodiments, the cyclic dinucleotide of Formula (I), (I-1),(I-2), (Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig) has thestructure:

or an enantiomer, hydrate, solvate or pharmaceutically acceptable saltthereof.

The presence of a chiral center allows the compound to exist as one oftwo possible optical isomers ((R)- or (S)-enantiomer) or as a racemicmixture of both. Where substituents are present that may be attached atdifferent positions in the molecule, all regioisomers and mixtures ofregioisomers formed are included within the scope of the Formula (I)described by this invention.

A compound of the disclosure, e.g, a compound of Formula (I), (I-1),(I-2), (Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig) can beshown in a number of equivalent depictions. For example, a compound ofFormula (Ia-2) is typically depicted herein as shown below:

The above compound of Formula (Ia-2) is equivalent to a compound ofFormula (Ia-2) as depicted below:

Further, each of the previous depictions are equivalent to the belowdepiction of a compound of Formula (Ia-2):

IV. Compositions

In certain embodiments, the present disclosure provides a pharmaceuticalcomposition comprising a compound of the present disclosure (e.g. acompound of Formula (I), (I-1), (I-2), (Ia-1), (Ia-2), (Ib), (Ic), (Id),(Ie), (If), and/or (Ig), or a pharmaceutically acceptable salt thereof,and a pharmaceutically acceptable excipient.

In certain embodiments, the pharmaceutical composition comprises one ormore additional therapeutic agent, as more fully set forth below.

Pharmaceutical compositions comprising the compounds disclosed herein,or pharmaceutically acceptable salts thereof, may be prepared with oneor more pharmaceutically acceptable excipients which may be selected inaccord with ordinary practice. Tablets may contain excipients includingglidants, fillers, binders and the like. Aqueous compositions may beprepared in sterile form, and when intended for delivery by other thanoral administration generally may be isotonic. All compositions mayoptionally contain excipients such as those set forth in the Rowe et al,Handbook of Pharmaceutical Excipients, 6^(th) edition, AmericanPharmacists Association, 2009. Excipients can include ascorbic acid andother antioxidants, chelating agents such as EDTA, carbohydrates such asdextrin, hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearicacid and the like. In certain embodiments, the composition is providedas a solid dosage form, including a solid oral dosage form.

The compositions include those suitable for various administrationroutes, including oral administration. The compositions may be presentedin unit dosage form and may be prepared by any of the methods well knownin the art of pharmacy. Such methods include the step of bringing intoassociation the active ingredient (e.g., a compound of the presentdisclosure or a pharmaceutical salt thereof) with one or morepharmaceutically acceptable excipients. The compositions may be preparedby uniformly and intimately bringing into association the activeingredient with liquid excipients or finely divided solid excipients orboth, and then, if necessary, shaping the product. Techniques andformulations generally are found in Remington: The Science and Practiceof Pharmacy, 21^(st) Edition, Lippincott Wiliams and Wilkins,Philadelphia, Pa., 2006.

Compositions described herein that are suitable for oral administrationmay be presented as discrete units (a unit dosage form) including butnot limited to capsules, cachets or tablets each containing apredetermined amount of the active ingredient. In one embodiment, thepharmaceutical composition is a tablet.

Pharmaceutical compositions disclosed herein comprise one or morecompounds disclosed herein, or a pharmaceutically acceptable saltthereof, together with a pharmaceutically acceptable excipient andoptionally other therapeutic agents. Pharmaceutical compositionscontaining the active ingredient may be in any form suitable for theintended method of administration. When used for oral use for example,tablets, troches, lozenges, aqueous or oil suspensions, dispersiblepowders or granules, emulsions, hard or soft capsules, syrups or elixirsmay be prepared. Compositions intended for oral use may be preparedaccording to any method known to the art for the manufacture ofpharmaceutical compositions and such compositions may contain one ormore excipients including sweetening agents, flavoring agents, coloringagents and preserving agents, in order to provide a palatablepreparation. Tablets containing the active ingredient in admixture withnon-toxic pharmaceutically acceptable excipients which are suitable formanufacture of tablets are acceptable. These excipients may be, forexample, inert diluents, such as calcium or sodium carbonate, lactose,lactose monohydrate, croscarmellose sodium, povidone, calcium or sodiumphosphate; granulating and disintegrating agents, such as maize starch,or alginic acid; binding agents, such as cellulose, microcrystallinecellulose, starch, gelatin or acacia; and lubricating agents, such asmagnesium stearate, stearic acid or talc. Tablets may be uncoated or maybe coated by known techniques including microencapsulation to delaydisintegration and adsorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearatealone or with a wax may be employed.

The amount of active ingredient that may be combined with the inactiveingredients to produce a dosage form may vary depending upon theintended treatment subject and the particular mode of administration.For example, in some embodiments, a dosage form for oral administrationto humans may contain approximately 1 to 1000 mg of active materialformulated with an appropriate and convenient amount of apharmaceutically acceptable excipient. In certain embodiments, thepharmaceutically acceptable excipient varies from about 5 to about 95%of the total compositions (weight:weight).

In certain embodiments, a composition comprising a compound of thepresent disclosure (e.g. a compound of Formula (I), (I-1), (I-2),(Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If), and/or (Ig)), or apharmaceutically acceptable salt thereof in one variation does notcontain an agent that affects the rate at which the active ingredient ismetabolized. Thus, it is understood that compositions comprising acompound of the present disclosure in one aspect do not comprise anagent that would affect (e.g., slow, hinder or retard) the metabolism ofa compound of the present disclosure or any other active ingredientadministered separately, sequentially or simultaneously with a compoundof the present disclosure. It is also understood that any of themethods, kits, articles of manufacture and the like detailed herein inone aspect do not comprise an agent that would affect (e.g., slow,hinder or retard) the metabolism of a compound of the present disclosureor any other active ingredient administered separately, sequentially orsimultaneously with a compound of the present disclosure.

The invention further includes a pharmaceutical composition as describedabove for use in modulating STING protein activity, to induceSTING-dependent production of type I interferons, cytokines orchemokines.

The invention further includes a pharmaceutical composition as describedabove for use in treating or preventing viral infection, infectioncaused by hepatitis B virus, by HIV, hyperproliferative disease orcancer.

The invention further includes a compound of the present disclosure foradministration as a single active ingredient of a pharmaceuticallyacceptable composition which can be prepared by conventional methodsknown in the art, for example by binding the active ingredient to apharmaceutically acceptable, therapeutically inert organic and/orinorganic carrier or excipient, or by mixing therewith.

Another possibility is the use of a compound of the present disclosureas a second or other active ingredient having a synergistic effect withother active ingredients in known drugs, or administration of thecompound of the present disclosure together with such drugs.

The compound of the present disclosure may also be used in the form of aprodrug or other suitably modified form which releases the activeingredient in vivo.

V. Methods

In one embodiment, provided herein is a method of treating a disease ordisorder, comprising administering to a human or animal in need thereofa therapeutically effective amount of a compound of the presentdisclosure or an enantiomer, pharmaceutically acceptable salt, hydrate,or solvate thereof.

Also provided is a method of modulating, e.g., increasing, the activityof STING protein, comprising administering a therapeutically effectiveamount of a compound of the present disclosure or an enantiomer,pharmaceutically acceptable salt, hydrate, or solvate thereof.

The Stimulator of interferon genes (STING) adaptor protein, also knownas STING, STING protein, transmembrane protein 173 (TMEM173), MPYS,mediator of IRF3 activation (MITA), or endoplasmic reticulum interferonstimulator (ERIS), is a protein that in humans is encoded by the TMEM173gene (UniProt code Q86WV6; NCBI Reference Sequences: NP_938023.1(isoform 1) and NP_001288667 (isoform 2)). STING adaptor protein isbelieved to function as both a direct cytosolic DNA sensor (CDS) and anadaptor protein in Type I interferon signaling through differentmolecular mechanisms. STING adaptor protein has been shown to activatedownstream transcription factors STAT6 and IRF3 through TBK1, and NF-κBthrough IKKβ, which can effect an antiviral response or innate immuneresponse against an intracellular pathogen. STING adaptor protein playsa role in innate immunity by inducing type I interferon production whencells are infected with intracellular pathogens, such as viruses,mycobacteria and intracellular parasites. Type I interferon, mediated bySTING adaptor protein, protects infected cells and nearby cells fromlocal infection by autocrine and paracrine signaling.

Further provided is a method of preventing or treating a disease orcondition responsive to the modulation of STING adaptor protein,comprising administering to a human or animal in need thereof atherapeutically effective amount of a compound of the presentdisclosure, or an enantiomer, pharmaceutically acceptable salt, hydrate,or solvate thereof.

Further provided is a method of inducing a STING adaptorprotein-dependent type I interferon, cytokine or chemokine in a human oranimal, comprising administering a therapeutically effective amount of acompound of the present disclosure, or an enantiomer, pharmaceuticallyacceptable salt, hydrate, or solvate thereof.

Activation of STING adaptor protein in turn activates protein kinaseTBK1, which subsequently activates downstream transcription factorsNF-κB and IRF-3. Activation of STING adaptor protein ultimately isbelieved to result in the release of type I and III interferons as wellas a variety of cytokines and chemokines such as IL-6, TNF-α and INF-γ.

Accordingly, induction of a STING adaptor protein-dependent type Iinterferon, cytokine or chemokine in a human or animal results in theactivation of one or more of NF-κB, IRF-3, a type I interferon, a typeIII interferon, IL-6, TNF-α, and INF-γ in said human or animal.

Further provided is a method of treating or preventing viral infection,e.g., infection by hepatitis B or HIV, comprising administering to ahuman or animal in need thereof a therapeutically effective amount of acompound of the present disclosure, or an enantiomer, pharmaceuticallyacceptable salt, hydrate, or solvate thereof.

Viral infections that can be treated or prevented by the methods of thepresent disclosure can be any infection caused by a virus, e.g., a virusfrom the Hepadnaviridae family of viruses, e.g., hepatitis B; or anyretrovirus, e.g., an alpharetrovirus, such as Rous sarcoma virus; abetaretrovirus, such as simian retrovirus; a deltaretrovirus, such asbovine leukemia virus or human T-lymphotrophic virus (HTLV) includivindHTLV-1, HTLV-2, and HTLV-3; a gammaretrovirus, such as murine leukemiavirus or feline leukemia virus; or a lentivirus, such as humanimmunodeficiency virus (HIV) including HIV-1 and HIV-2, simianimmunodeficiency virus, equine infectious anemia virus, bovineimmunodeficiency virus, rabbit endogenous lentivirus type K (RELIK), orfeline immunodeficiency virus.

Further provided is a method of treating or preventing ahyperproliferative disease or cancer, comprising administering to ahuman or animal in need thereof a therapeutically effective amount of acompound of the present disclosure, or an enantiomer, pharmaceuticallyacceptable salt, hydrate, or solvate thereof.

Hyperproliferative diseases include diseases caused by excessive growthof non-cancer cells. Such conditions include but are not limited topsoriasis, actinic keratoses, and seborrheic keratoses, warts, keloids,and eczema.

Cancers that can be treated or prevented by the methods of thedisclosure include solid tumors and lymphomas, including but not limitedto adrenal cancer, bladder cancer, bone cancer, brain cancer, breastcancer, colon cancer, colorectal cancer, eye cancer, head-and-neckcancer, kidney cancer such as renal cell carcinoma, liver cancer, lungcancer such as non-small cell lung cancer, ovarian cancer, pancreaticcancer, prostate cancer, skin cancer such as squamous cell carcinoma andmelanoma, thyroid cancer, uterine cancer, vaginal cancer, and myelomasuch as multiple myeloma. The cancer can be naïve, or relapsed and/orrefractory.

Further provided is a method of enhancing the efficacy of a vaccine,comprising administering to a human or animal in need thereof atherapeutically effective amount of a compound of the presentdisclosure, or an enantiomer, pharmaceutically acceptable salt, hydrate,or solvate thereof.

The invention includes a compound of the present disclosure, or anenantiomer, pharmaceutically acceptable salt, hydrate, or solvatethereof for use as a medicament in a human or animal.

The invention further includes a compound of the present disclosure, oran enantiomer, pharmaceutically acceptable salt, hydrate, or solvatethereof for use in modulating the activity of STING protein.

The invention further includes a compound of the present disclosure, oran enantiomer, pharmaceutically acceptable salt, hydrate, or solvatethereof for use in the prevention or treatment of a disease or conditionin a human or animal responsive to the modulation of the STING protein.

The invention further includes a compound of the present disclosure, oran enantiomer, pharmaceutically acceptable salt, hydrate, or solvatethereof alone or in combination with one or more therapeutically activesubstances, for use in STING dependent induction of a type I interferon,cytokine or chemokine in a human or animal.

The invention further includes a compound of the present disclosure, oran enantiomer, pharmaceutically acceptable salt, hydrate, or solvatethereof, alone or in combination with one or more therapeutically activeagents for use in the treatment or prevention of viral infection in ahuman or animal.

The invention further includes a compound of the present disclosure, oran enantiomer, pharmaceutically acceptable salt, hydrate, or solvatethereof, alone or in combination with one or more therapeutically activesubstances, for use in the treatment or prevention of infection causedby hepatitis B virus or HIV in a human or animal.

The invention further includes a compound of the present disclosure, oran enantiomer, pharmaceutically acceptable salt, hydrate, or solvatethereof alone or in combination with one or more therapeutically activeagents, for use in the treatment or prevention of a hyperproliferativedisease or cancer in a human or animal.

The invention further includes a compound of the present disclosure, oran enantiomer, pharmaceutically acceptable salt, hydrate, or solvatethereof for use in enhancing vaccine efficacy in a human or animal.

The invention further includes a pharmaceutical composition for use inmodulating STING protein activity, to induce STING-dependent productionof a type I interferon, cytokine or chemokine in a human or animal.

The invention further includes a pharmaceutical composition for use intreating or preventing viral infection, infection caused by hepatitis Bvirus, by HIV, hyperproliferative disease or cancer in a human oranimal.

The invention further includes the use of a compound of the presentdisclosure, or an enantiomer, pharmaceutically acceptable salt, hydrate,or solvate thereof for the production of a medicament for the treatmentor prevention of infection caused by hepatitis B virus, by HIV, ofhyperproliferative disease or cancer.

VI. Administration

The compounds of the present disclosure (also referred to herein as theactive ingredients), can be administered by any route appropriate to thecondition to be treated. Suitable routes include oral, rectal, nasal,topical (including buccal and sublingual), transdermal, vaginal andparenteral (including subcutaneous, intramuscular, intravenous,intradermal, intrathecal and epidural), and the like. It will beappreciated that the preferred route may vary with for example thecondition of the recipient. An advantage of certain compounds disclosedherein is that they are orally bioavailable and can be dosed orally.

A compound of the present disclosure, may be administered to anindividual in accordance with an effective dosing regimen for a desiredperiod of time or duration, such as at least about one month, at leastabout 2 months, at least about 3 months, at least about 6 months, or atleast about 12 months or longer. In one variation, the compound isadministered on a daily or intermittent schedule for the duration of theindividual's life.

The dosage or dosing frequency of a compound of the present disclosuremay be adjusted over the course of the treatment, based on the judgmentof the administering physician.

The compound may be administered to an individual (e.g., a human) in aneffective amount. In certain embodiments, the compound is administeredonce daily.

The compound can be administered by any useful route and means, such asby oral or parenteral (e.g., intravenous) administration.Therapeutically effective amounts of the compound may include from about0.00001 mg/kg body weight per day to about 10 mg/kg body weight per day,such as from about 0.0001 mg/kg body weight per day to about 10 mg/kgbody weight per day, or such as from about 0.001 mg/kg body weight perday to about 1 mg/kg body weight per day, or such as from about 0.01mg/kg body weight per day to about 1 mg/kg body weight per day, or suchas from about 0.05 mg/kg body weight per day to about 0.5 mg/kg bodyweight per day, or such as from about 0.3 mg to about 30 mg per day, orsuch as from about 30 mg to about 300 mg per day.

A compound of the present disclosure may be combined with one or moreadditional therapeutic agents in any dosage amount of the compound ofthe present disclosure (e.g., from 1 mg to 1000 mg of compound).Therapeutically effective amounts may include from about 1 mg per doseto about 1000 mg per dose, such as from about 50 mg per dose to about500 mg per dose, or such as from about 100 mg per dose to about 400 mgper dose, or such as from about 150 mg per dose to about 350 mg perdose, or such as from about 200 mg per dose to about 300 mg per dose.Other therapeutically effective amounts of the compound of the presentdisclosure are about 100, 125, 150, 175, 200, 225, 250, 275, 300, 325,350, 375, 400, 425, 450, 475, or about 500 mg per dose. Othertherapeutically effective amounts of the compound of the presentdisclosure are about 100 mg per dose, or about 125, 150, 175, 200, 225,250, 275, 300, 350, 400, 450, or about 500 mg per dose. A single dosecan be administered hourly, daily, or weekly. For example, a single dosecan be administered once every 1 hour, 2, 3, 4, 6, 8, 12, 16 or onceevery 24 hours. A single dose can also be administered once every 1 day,2, 3, 4, 5, 6, or once every 7 days. A single dose can also beadministered once every 1 week, 2, 3, or once every 4 weeks. In certainembodiments, a single dose can be administered once every week. A singledose can also be administered once every month.

Kits that comprise a compound of the present disclosure, or anenantiomer, pharmaceutically acceptable salt, hydrate, or solvatethereof, or a pharmaceutical composition containing any of the above,are also included in the present invention.

In one embodiment, kits comprising a compound disclosed herein, or apharmaceutically acceptable salt thereof, in combination with one ormore (e.g., one, two, three, four, one or two, or one to three, or oneto four) additional therapeutic agents are provided.

VII. Combination Therapy

In certain embodiments, a method for treating or preventing aninfectious disease, a viral infection, hepatitis B infection, HIVinfection, cancer, or a hyperproliferative disease in a human having orat risk of having the disease is provided, comprising administering tothe human a therapeutically effective amount of a compound of thepresent disclosure, e.g., a compound of Formula (I), (I-1), (I-2),(Ia-1), (Ia-2), (Ib), (Ic), (Id), (Ie), (If) and/or (Ig), or apharmaceutically acceptable salt thereof, in combination with atherapeutically effective amount of one or more (e.g., one, two, three,four, one or two, one to three, or one to four) additional therapeuticagents. In one embodiment, a method for treating an infectious disease,a viral infection, hepatitis B infection, HIV infection, cancer, or ahyperproliferative disease in a human having or at risk of having thedisease is provided, comprising administering to the human atherapeutically effective amount of a compound disclosed herein, or apharmaceutically acceptable salt thereof, in combination with atherapeutically effective amount of one or more (e.g., one, two, three,four, one or two, one to three, or one to four) additional therapeuticagents.

In certain embodiments, the present disclosure provides a method fortreating a viral infection, comprising administering to a subject inneed thereof a therapeutically effective amount of a compound disclosedherein or a pharmaceutically acceptable salt thereof, in combinationwith a therapeutically effective amount of one or more (e.g., one, two,three, four, one or two, one to three, or one to four) additionaltherapeutic agents which are suitable for treating the viral infection.In some embodiments, the viral infection is a hepatitis B infection. Insome embodiments, the viral infection is a HIV infection.

In certain embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with one, two,three, four, or more additional therapeutic agents. In certainembodiments, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with two additional therapeuticagents. In other embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with threeadditional therapeutic agents. In further embodiments, a compounddisclosed herein, or a pharmaceutically acceptable salt thereof, iscombined with four additional therapeutic agents. The one, two, three,four, or more additional therapeutic agents can be different therapeuticagents selected from the same class of therapeutic agents, and/or theycan be selected from different classes of therapeutic agents.

Administration of Combination Therapy

In certain embodiments, a compound disclosed herein is administered withone or more additional therapeutic agents. Co-administration of acompound disclosed herein with one or more additional therapeutic agentsgenerally refers to simultaneous or sequential administration of acompound disclosed herein and one or more additional therapeutic agents,such that therapeutically effective amounts of the compound disclosedherein and the one or more additional therapeutic agents are bothpresent in the body of the subject. When administered sequentially, thecombination may be administered in two or more administrations.

Co-administration of a compound disclosed herein with one or moreadditional therapeutic agents generally refers to simultaneous orsequential administration of a compound disclosed herein and one or moreadditional therapeutic agents, such that therapeutically effectiveamounts of each agent are present in the body of the patient.

In certain embodiments, a compound of the present disclosure may becombined with one or more (e.g., one, two, three, four, one or two, oneto three, or one to four) additional therapeutic agents in any dosageamount of the compound (e.g., from 10 mg to 1000 mg of compound).

Co-administration includes administration of unit dosages of thecompounds disclosed herein before or after administration of unitdosages of one or more additional therapeutic agents. The compounddisclosed herein may be administered within seconds, minutes, or hoursof the administration of one or more additional therapeutic agents. Forexample, in some embodiments, a unit dose of a compound disclosed hereinis administered first, followed within seconds or minutes byadministration of a unit dose of one or more additional therapeuticagents. Alternatively, in other embodiments, a unit dose of one or moreadditional therapeutic agents is administered first, followed byadministration of a unit dose of a compound disclosed herein withinseconds or minutes. In some embodiments, a unit dose of a compounddisclosed herein is administered first, followed, after a period ofhours (e.g., 1-12 hours), by administration of a unit dose of one ormore additional therapeutic agents. In other embodiments, a unit dose ofone or more additional therapeutic agents is administered first,followed, after a period of hours (e.g., 1-12 hours), by administrationof a unit dose of a compound disclosed herein.

In certain embodiments, a compound disclosed herein is combined with oneor more additional therapeutic agents in a unitary dosage form forsimultaneous administration to a subject, for example as a solid dosageform for oral administration.

In certain embodiments a compound of the present disclosure isformulated as a tablet, which may optionally contain one or more othercompounds useful for treating the disease being treated. In certainembodiments, the tablet can contain another active ingredient fortreating a viral disease, e.g., hepatitis B virus or HIV.

In certain embodiments, such tablets are suitable for once daily dosing.

In one embodiment, pharmaceutical compositions comprising a compounddisclosed herein, or a pharmaceutically acceptable salt thereof, incombination with one or more (e.g., one, two, three, one or two, or oneto three) additional therapeutic agents, and a pharmaceuticallyacceptable carrier, diluent, or excipient are provided.

In one embodiment, kits comprising a compound disclosed herein, or apharmaceutically acceptable salt thereof, in combination with one ormore (e.g., one, two, three, four, one or two, or one to three, or oneto four) additional therapeutic agents are provided.

Viral Combination Therapy

The compounds described herein may be used or combined with one or moreof a antiviral agents including abacavir, aciclovir, adefovir,amantadine, amprenavir, arbidol, atazanavir, artipla, brivudine,cidofovir, combivir, edoxudine, efavirenz, emtricitabine, enfuvirtide,entecavir, fomvirsen, fosamprenavir, foscarnet, fosfonet, ganciclovir,gardasil, ibacitabine, immunovir, idoxuridine, imiquimod, indinavir,inosine, integrase inhibitors, interferons, including interferon typeIII, interferon type II, interferon type I, lamivudine, lopinavir,loviride, MK-0518, maraviroc, moroxydine, nelfinavir, nevirapine,nexavir, nucleoside analogues, oseltamivir, penciclovir, peramivir,pleconaril, podophyllotoxin, protease inhibitors, reverse transcriptaseinhibitors, ribavirin, rimantadine, ritonavir, saquinavir, stavudine,tenofovir, tenofovir disoproxil, tipranavir, trifluridine, trizivir,tromantadine, truvada, valganciclovir, vicriviroc, vidarabine,viramidine, zalcitabine, zanamivir, zidovudine, and combinationsthereof.

In certain embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with 5-30 mgtenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, ortenofovir alafenamide. In certain embodiments, a compound disclosedherein, or a pharmaceutically acceptable salt thereof, is combined with5-10; 5-15; 5-20; 5-25; 25-30; 20-30; 15-30; or 10-30 mg tenofoviralafenamide fumarate, tenofovir alafenamide hemifumarate, or tenofoviralafenamide. In certain embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with 10 mgtenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, ortenofovir alafenamide. In certain embodiments, a compound disclosedherein, or a pharmaceutically acceptable salt thereof, is combined with25 mg tenofovir alafenamide fumarate, tenofovir alafenamidehemifumarate, or tenofovir alafenamide. A compound as disclosed herein(e.g., a compound of Formula (I)) may be combined with the agentsprovided herein in any dosage amount of the compound (e.g., from 50 mgto 500 mg of compound) the same as if each combination of dosages werespecifically and individually listed.

In certain embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with 100-400 mgtenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, ortenofovir disoproxil. In certain embodiments, a compound disclosedherein, or a pharmaceutically acceptable salt thereof, is combined with100-150; 100-200, 100-250; 100-300; 100-350; 150-200; 150-250; 150-300;150-350; 150-400; 200-250; 200-300; 200-350; 200-400; 250-350; 250-400;350-400 or 300-400 mg tenofovir disoproxil fumarate, tenofovirdisoproxil hemifumarate, or tenofovir disoproxil. In certainembodiments, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with 300 mg tenofovir disoproxilfumarate, tenofovir disoproxil hemifumarate, or tenofovir disoproxil. Incertain embodiments, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with 250 mg tenofovir disoproxilfumarate, tenofovir disoproxil hemifumarate, or tenofovir disoproxil. Incertain embodiments, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with 150 mg tenofovir disoproxilfumarate, tenofovir disoproxil hemifumarate, or tenofovir disoproxil. Acompound as disclosed herein (e.g., a compound of Formula (I)) may becombined with the agents provided herein in any dosage amount of thecompound (e.g., from 50 mg to 500 mg of compound) the same as if eachcombination of dosages were specifically and individually listed.

HIV Combination Therapy

In certain embodiments, a method for treating or preventing an HIVinfection in a human or animal having or at risk of having the infectionis provided, comprising administering to the human or animal atherapeutically effective amount of a compound disclosed herein, or apharmaceutically acceptable salt thereof, in combination with atherapeutically effective amount of one or more (e.g., one, two, three,one or two, or one to three) additional therapeutic agents. In oneembodiment, a method for treating an HIV infection in a human or animalhaving or at risk of having the infection is provided, comprisingadministering to the human or animal a therapeutically effective amountof a compound disclosed herein, or a pharmaceutically acceptable saltthereof, in combination with a therapeutically effective amount of oneor more (e.g., one, two, three, one or two, or one to three) additionaltherapeutic agents.

In certain embodiments, the present disclosure provides a method fortreating an HIV infection, comprising administering to a subject in needthereof a therapeutically effective amount of a compound disclosedherein, or a pharmaceutically acceptable salt thereof, in combinationwith a therapeutically effective amount of one or more additionaltherapeutic agents which are suitable for treating an HIV infection.

In certain embodiments, the compounds disclosed herein are formulated asa tablet, which may optionally contain one or more other compoundsuseful for treating HIV. In certain embodiments, the tablet can containanother active ingredient for treating HIV, such as HIV proteaseinhibitors, HIV non-nucleoside or non-nucleotide inhibitors of reversetranscriptase, HIV nucleoside or nucleotide inhibitors of reversetranscriptase, HIV integrase inhibitors, HIV non-catalytic site (orallosteric) integrase inhibitors, pharmacokinetic enhancers, andcombinations thereof.

In certain embodiments, such tablets are suitable for once daily dosing.

In the above embodiments, the additional therapeutic agent may be ananti-HIV agent. In some embodiments, the additional therapeutic agent isselected from the group consisting of HIV combination drugs, HIVprotease inhibitors, HIV non-nucleoside or non-nucleotide inhibitors ofreverse transcriptase, HIV nucleoside or nucleotide inhibitors ofreverse transcriptase, HIV integrase inhibitors, HIV non-catalytic site(or allosteric) integrase inhibitors, HIV entry inhibitors, HIVmaturation inhibitors, immunomodulators, immunotherapeutic agents,antibody-drug conjugates, gene modifiers, gene editors (such asCRISPR/Cas9, zinc finger nucleases, homing nucleases, syntheticnucleases, TALENs), cell therapies (such as chimeric antigen receptorT-cell, CAR-T, and engineered T cell receptors, TCR-T), latencyreversing agents, compounds that target the HIV capsid (including capsidinhibitors), immune-based therapies, phosphatidylinositol 3-kinase(PI3K) inhibitors, alpha-4/beta-7 antagonists, HIV antibodies,bispecific antibodies and “antibody-like” therapeutic proteins, HIV p17matrix protein inhibitors, IL-13 antagonists, peptidyl-prolyl cis-transisomerase A modulators, protein disulfide isomerase inhibitors,complement C5a receptor antagonists, DNA methyltransferase inhibitor,HIV vif gene modulators, Vif dimerization antagonists, HIV-1 viralinfectivity factor inhibitors, TAT protein inhibitors, HIV-1 Nefmodulators, Hck tyrosine kinase modulators, mixed lineage kinase-3(MLK-3) inhibitors, HIV-1 splicing inhibitors, Rev protein inhibitors,integrin antagonists, nucleoprotein inhibitors, splicing factormodulators, COMM domain containing protein 1 modulators, HIVribonuclease H inhibitors, retrocyclin modulators, CDK-9 inhibitors,dendritic ICAM-3 grabbing nonintegrin 1 inhibitors, HIV GAG proteininhibitors, HIV POL protein inhibitors, Complement Factor H modulators,ubiquitin ligase inhibitors, deoxycytidine kinase inhibitors, cyclindependent kinase inhibitors, proprotein convertase PC9 stimulators, ATPdependent RNA helicase DDX3X inhibitors, reverse transcriptase primingcomplex inhibitors, G6PD and NADH-oxidase inhibitors, pharmacokineticenhancers, HIV gene therapy, HIV vaccines, and other HIV therapeuticagents, and combinations thereof.

In some embodiments, the additional therapeutic agent is selected fromthe group consisting of combination drugs for HIV, other drugs fortreating HIV, HIV protease inhibitors, HIV reverse transcriptaseinhibitors, HIV integrase inhibitors, HIV non-catalytic site (orallosteric) integrase inhibitors, HIV entry (fusion) inhibitors, HIVmaturation inhibitors, latency reversing agents, capsid inhibitors,immune-based therapies, PI3K inhibitors, HIV antibodies, and bispecificantibodies, and “antibody-like” therapeutic proteins, and combinationsthereof.

HIV Combination Drugs

Examples of combination drugs include ATRIPLA® (efavirenz, tenofovirdisoproxil fumarate, and emtricitabine); COMPLERA® (EVIPLERA®;rilpivirine, tenofovir disoproxil fumarate, and emtricitabine);STRIBILD® (elvitegravir, cobicistat, tenofovir disoproxil fumarate, andemtricitabine); TRUVADA® (tenofovir disoproxil fumarate andemtricitabine; TDF+FTC); DESCOVY® (tenofovir alafenamide andemtricitabine); ODEFSEY® (tenofovir alafenamide, emtricitabine, andrilpivirine); GENVOYA® (tenofovir alafenamide, emtricitabine,cobicistat, and elvitegravir); BIKTARVY® (bictegravir, emtricitabine,tenofovir alafenamide); darunavir, tenofovir alafenamide hemifumarate,emtricitabine, and cobicistat; efavirenz, lamivudine, and tenofovirdisoproxil fumarate; lamivudine and tenofovir disoproxil fumarate;tenofovir and lamivudine; tenofovir alafenamide and emtricitabine;tenofovir alafenamide hemifumarate and emtricitabine; tenofoviralafenamide hemifumarate, emtricitabine, and rilpivirine; tenofoviralafenamide hemifumarate, emtricitabine, cobicistat, and elvitegravir;COMBIVIR® (zidovudine and lamivudine; AZT+3TC); EPZICOM® (LIVEXA®;abacavir sulfate and lamivudine; ABC+3TC); KALETRA® (ALUVIA®; lopinavirand ritonavir); TRIUMEQ® (dolutegravir, abacavir, and lamivudine);TRIZIVIR® (abacavir sulfate, zidovudine, and lamivudine; ABC+AZT+3TC);atazanavir and cobicistat; atazanavir sulfate and cobicistat; atazanavirsulfate and ritonavir; darunavir and cobicistat; dolutegravir andrilpivirine; dolutegravir and rilpivirine hydrochloride; dolutegravir,abacavir sulfate, and lamivudine; lamivudine, nevirapine, andzidovudine; raltegravir and lamivudine; doravirine, lamivudine, andtenofovir disoproxil fumarate; doravirine, lamivudine, and tenofovirdisoproxil; dolutegravir+lamivudine, lamivudine+abacavir+zidovudine,lamivudine+abacavir, lamivudine+tenofovir disoproxil fumarate,lamivudine+zidovudine+nevirapine, lopinavir+ritonavir,lopinavir+ritonavir+abacavir+lamivudine,lopinavir+ritonavir+zidovudine+lamivudine, tenofovir+lamivudine, andtenofovir disoproxil fumarate+emtricitabine+rilpivirine hydrochloride,lopinavir, ritonavir, zidovudine and lamivudine; Vacc-4x and romidepsin;and APH-0812.

HIV Protease Inhibitors

Examples of HIV protease inhibitors include amprenavir, atazanavir,brecanavir, darunavir, fosamprenavir, fosamprenavir calcium, indinavir,indinavir sulfate, lopinavir, nelfinavir, nelfinavir mesylate,ritonavir, saquinavir, saquinavir mesylate, tipranavir, DG-17, TMB-657(PPL-100), T-169, BL-008, and TMC-310911.

HIV Reverse Transcriptase Inhibitors

Examples of HIV non-nucleoside or non-nucleotide inhibitors of reversetranscriptase include dapivirine, delavirdine, delavirdine mesylate,doravirine, efavirenz, etravirine, lentinan, nevirapine, rilpivirine,ACC-007, AIC-292, KM-023, PC-1005, and VM-1500.

Examples of HIV nucleoside or nucleotide inhibitors of reversetranscriptase include adefovir, adefovir dipivoxil, azvudine,emtricitabine, tenofovir, tenofovir alafenamide, tenofovir alafenamidefumarate, tenofovir alafenamide hemifumarate, tenofovir disoproxil,tenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, VIDEX®and VIDEX EC® (didanosine, ddl), abacavir, abacavir sulfate, alovudine,apricitabine, censavudine, didanosine, elvucitabine, festinavir,fosalvudine tidoxil, CMX-157, dapivirine, doravirine, etravirine,OCR-5753, tenofovir disoproxil orotate, fozivudine tidoxil, lamivudine,phosphazid, stavudine, zalcitabine, zidovudine, GS-9131, GS-9148,MK-8504 and KP-1461.

HIV Integrase Inhibitors

Examples of HIV integrase inhibitors include elvitegravir, curcumin,derivatives of curcumin, chicoric acid, derivatives of chicoric acid,3,5-dicaffeoylquinic acid, derivatives of 3,5-dicaffeoylquinic acid,aurintricarboxylic acid, derivatives of aurintricarboxylic acid, caffeicacid phenethyl ester, derivatives of caffeic acid phenethyl ester,tyrphostin, derivatives of tyrphostin, quercetin, derivatives ofquercetin, raltegravir, dolutegravir, JTK-351, bictegravir, AVX-15567,cabotegravir (long-acting injectable), diketo quinolin-4-1 derivatives,integrase-LEDGF inhibitor, ledgins, M-522, M-532, NSC-310217,NSC-371056, NSC-48240, NSC-642710, NSC-699171, NSC-699172, NSC-699173,NSC-699174, stilbenedisulfonic acid, T-169 and cabotegravir.

Examples of HIV non-catalytic site, or allosteric, integrase inhibitors(NCINI) include CX-05045, CX-05168, and CX-14442.

HIV Entry Inhibitors

Examples of HIV entry (fusion) inhibitors include cenicriviroc, CCR5inhibitors, gp41 inhibitors, CD4 attachment inhibitors, gp120inhibitors, and CXCR4 inhibitors. Examples of CCR5 inhibitors includeaplaviroc, vicriviroc, maraviroc, cenicriviroc, PRO-140, adaptavir(RAP-101), nifeviroc (TD-0232), anti-GP120/CD4 or CCR5 bispecificantibodies, B-07, MB-66, polypeptide C25P, TD-0680, and vMIP (Haimipu).

Examples of gp41 inhibitors include albuvirtide, enfuvirtide,BMS-986197, enfuvirtide biobetter, enfuvirtide biosimilar, HIV-1 fusioninhibitors (P26-Bapc), ITV-1, ITV-2, ITV-3, ITV-4, PIE-12 trimer andsifuvirtide.

Examples of CD4 attachment inhibitors include ibalizumab and CADAanalogs

Examples of gp120 inhibitors include Radha-108 (receptol) 3B3-PE38,BanLec, bentonite-based nanomedicine, fostemsavir tromethamine,IQP-0831, and BMS-663068.

Examples of CXCR4 inhibitors include plerixafor, ALT-1188, N15 peptide,and vMIP (Haimipu).

HIV Maturation Inhibitors

Examples of HIV maturation inhibitors include BMS-955176 andGSK-2838232.

Latency Reversing Agents

Examples of latency reversing agents include histone deacetylase (HDAC)inhibitors, proteasome inhibitors such as velcade, protein kinase C(PKC) activators, Smyd2 inhibitors, BET-bromodomain 4 (BRD4) inhibitors,ionomycin, PMA, SAHA (suberanilohydroxamic acid, or suberoyl, anilide,and hydroxamic acid), AM-0015, ALT-803, NIZ-985, NKTR-255, IL-15modulating antibodies, JQ1, disulfiram, amphotericin B, and ubiquitininhibitors such as largazole analogs, and GSK-343.

Examples of HDAC inhibitors include romidepsin, vorinostat, andpanobinostat.

Examples of PKC activators include indolactam, prostratin, ingenol B,and DAG-lactones.

Capsid Inhibitors

Examples of capsid inhibitors include capsid polymerization inhibitorsor capsid disrupting compounds, HIV nucleocapsid p7 (NCp7) inhibitorssuch as azodicarbonamide, HIV p24 capsid protein inhibitors, AVI-621,AVI-101, AVI-201, AVI-301, and AVI-CAN1-15 series;

Immune-Based Therapies

Examples of immune-based therapies include toll-like receptorsmodulators such as TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9,TLR10, TLR11, TLR12, and TLR13; programmed cell death protein 1 (Pd-1)modulators; programmed death-ligand 1 (Pd-L1) modulators; IL-15modulators; DermaVir; interleukin-7; plaquenil (hydroxychloroquine);proleukin (aldesleukin, IL-2); interferon alfa; interferon alfa-2b;interferon alfa-n3; pegylated interferon alfa; interferon gamma;hydroxyurea; mycophenolate mofetil (MPA) and its ester derivativemycophenolate mofetil (MMF); ribavirin; rintatolimod, polymerpolyethyleneimine (PEI); gepon; rintatolimod; IL-12; WF-10; VGV-1;MOR-22; BMS-936559; CYT-107, interleukin-15/Fc fusion protein,normferon, peginterferon alfa-2a, peginterferon alfa-2b, recombinantinterleukin-15, RPI-MN, GS-9620, STING modulators, RIG-I modulators,NOD2 modulators, and IR-103.

Examples of TLR8 modulators include motolimod, resiquimod, 3M-051,3M-052, MCT-465, IMO-4200, VTX-763, VTX-1463 and those disclosed inUS20140045849 (Janssen), US20140073642 (Janssen), WO2014/056953(Janssen), WO2014/076221 (Janssen), WO2014/128189 (Janssen),US20140350031 (Janssen), WO2014/023813 (Janssen), US20080234251 (ArrayBiopharma), US20080306050 (Array Biopharma), US20100029585 (VentirxPharma), US20110092485 (Ventirx Pharma), US20110118235 (Ventirx Pharma),US20120082658 (Ventirx Pharma), US20120219615 (Ventirx Pharma),US20140066432 (Ventirx Pharma), US20140088085 (VentirxPharma),US20140275167 (Novira therapeutics), US20130251673 (Noviratherapeutics), U.S. Pat. No. 9,670,205 (Gilead Sciences Inc.),US20160289229 (Gilead Sciences Inc.), U.S. patent application Ser. No.15/692,161 (Gilead Sciences Inc.), and U.S. patent application Ser. No.15/692,093 (Gilead Sciences Inc.)

Phosphatidylinositol 3-Kinase (PI3K) Inhibitors

Examples of PI3K inhibitors include idelalisib, alpelisib, buparlisib,CAI orotate, copanlisib, duvelisib, gedatolisib, neratinib, panulisib,perifosine, pictilisib, pilaralisib, puquitinib mesylate, rigosertib,rigosertib sodium, sonolisib, taselisib, AMG-319, AZD-8186, BAY-1082439,CLR-1401, CLR-457, CUDC-907, DS-7423, EN-3342, GSK-2126458, GSK-2269577,GSK-2636771, INCB-040093, LY-3023414, MLN-1117, PQR-309, RG-7666,RP-6530, RV-1729, SAR-245409, SAR-260301, SF-1126, TGR-1202, UCB-5857,VS-5584, XL-765, and ZSTK-474.

Alpha-4/Beta-7 Antagonists

Examples of Integrin alpha-4/beta-7 antagonists include PTG-100,TRK-170, abrilumab, etrolizumab, carotegrast methyl, and vedolizumab.

HIV Antibodies, Bispecific Antibodies, and “Antibody-Like” TherapeuticProteins

Examples of HIV antibodies, bispecific antibodies, and “antibody-like”therapeutic proteins include DARTs®, DUOBODIES®, BITES®, XmAbs®,TandAbs®, Fab derivatives, bnABs (broadly neutralizing HIV-1antibodies), BMS-936559, TMB-360, and those targeting HIV gp120 or gp41,antibody-Recruiting Molecules targeting HIV, anti-CD63 monoclonalantibodies, anti-GB virus C antibodies, anti-GP120/CD4, CCR5 bispecificantibodies, anti-nef single domain antibodies, anti-Rev antibody,camelid derived anti-CD18 antibodies, camelid-derived anti-ICAM-1antibodies, DCVax-001, gp140 targeted antibodies, gp41-based HIVtherapeutic antibodies, human recombinant mAbs (PGT-121), ibalizumab,Immuglo, and MB-66.

Examples of those targeting HIV in such a manner include bavituximab,UB-421, C2F5, 2G12, C4E10, C2F5+C2G12+C4E10, 8ANC195, 3BNC117, 3BNC60,10-1074, PGT145, PGT121, PGT-151, PGT-133, MDX010 (ipilimumab), DH511,N6, VRC01 PGDM1400, A32, 7B2, 10E8, 10E8v4, CAP256-VRC26.25, DRVIA7,VRC-07-523, VRC-HIVMAB080-00-AB, VRC-HIVMAB060-00-AB, MGD-014 and VRC07.Example of HIV bispecific antibodies include MGD014.

Pharmacokinetic Enhancers

Examples of pharmacokinetic enhancers include cobicistat and ritonavir.

HIV Vaccines

Examples of HIV vaccines include peptide vaccines, recombinant subunitprotein vaccines, live vector vaccines, DNA vaccines, CD4-derivedpeptide vaccines, vaccine combinations, rgp120 (AIDSVAX), ALVAC HIV(vCP1521)/AIDSVAX B/E (gp120) (RV144), monomeric gp120 HIV-1 subtype Cvaccine, Remune, ITV-1, Contre Vir, Ad5-ENVA-48, DCVax-001 (CDX-2401),Vacc-4x, Vacc-C5, VAC-3S, multiclade DNA recombinant adenovirus-5(rAd5), Pennvax-G, Pennvax-GP, HIV-TriMix-mRNA vaccine, HIV-LAMP-vaxq,Ad35, Ad35-GRIN, NAcGM3/VSSP ISA-51, poly-ICLC adjuvanted vaccines,TatImmune, GTU-multiHIV (FIT-06), gp140[delta]V2.TV1+MF-59, rVSVIN HIV-1gag vaccine, SeV-Gag vaccine, AT-20, DNK-4, ad35-Grin/ENV, TBC-M4,HIVAX, HIVAX-2, NYVAC-HIV-PT1, NYVAC-HIV-PT4, DNA-HIV-PT123,rAAV1-PG9DP, GOVX-B11, GOVX-B21, TVI-HIV-1, Ad-4 (Ad4-env CladeC+Ad4-mGag), EN41-UGR7C, EN41-FPA2, PreVaxTat, AE-H, MYM-V101,CombiHIVvac, ADVAX, MYM-V201, MVA-CMDR, DNA-Ad5 gag/pol/nef/nev(HVTN505), MVATG-17401, ETV-01, CDX-1401, rcAD26.MOS1.HIV-Env,Ad26.Mod.HIV vaccine, AGS-004, AVX-101, AVX-201, PEP-6409, SAV-001,ThV-01, TL-01, TUTI-16, VGX-3300, IHV-001, and virus-like particlevaccines such as pseudovirion vaccine, CombiVICHvac, LFn-p24 B/C fusionvaccine, GTU-based DNA vaccine, HIV gag/pol/nef/env DNA vaccine,anti-TAT HIV vaccine, conjugate polypeptides vaccine, dendritic-cellvaccines, gag-based DNA vaccine, GI-2010, gp41 HIV-1 vaccine, HIVvaccine (PIKA adjuvant), I i-key/MHC class II epitope hybrid peptidevaccines, ITV-2, ITV-3, ITV-4, LIPO-5, multiclade Env vaccine, MVAvaccine, Pennvax-GP, pp71-deficient HCMV vector HIV gag vaccine,recombinant peptide vaccine (HIV infection), NCI, rgp160 HIV vaccine,RNActive HIV vaccine, SCB-703, Tat Oyi vaccine, TBC-M4, therapeutic HIVvaccine, UBI HIV gp120, Vacc-4x+romidepsin, variant gp120 polypeptidevaccine, rAd5 gag-pol env A/B/C vaccine, DNA.HTI and MVA.HTI.

Additional HIV Therapeutic Agents

Examples of additional HIV therapeutic agents include the compoundsdisclosed in WO 2004/096286 (Gilead Sciences), WO 2006/015261 (GileadSciences), WO 2006/110157 (Gilead Sciences), WO 2012/003497 (GileadSciences), WO 2012/003498 (Gilead Sciences), WO 2012/145728 (GileadSciences), WO 2013/006738 (Gilead Sciences), WO 2013/159064 (GileadSciences), WO 2014/100323 (Gilead Sciences), US 2013/0165489 (Universityof Pennsylvania), US 2014/0221378 (Japan Tobacco), US 2014/0221380(Japan Tobacco), WO 2009/062285 (Boehringer Ingelheim), WO 2010/130034(Boehringer Ingelheim), WO 2013/006792 (Pharma Resources), US20140221356 (Gilead Sciences), US 20100143301 (Gilead Sciences) and WO2013/091096 (Boehringer Ingelheim).

Examples of other drugs for treating HIV include acemannan, alisporivir,BanLec, deferiprone, Gamimune, metenkefalin, naltrexone, Prolastin, REP9, RPI-MN, VSSP, Hlviral, SB-728-T, 1,5-dicaffeoylquinic acid,rHIV7-shl-TAR-CCR5RZ, AAV-eCD4-Ig gene therapy, MazF gene therapy,BlockAide, ABX-464, AG-1105, APH-0812, BIT-225, CYT-107, HGTV-43,HPH-116, HS-10234, IMO-3100, IND-02, MK-1376, MK-8507, MK-8591, NOV-205,PA-1050040 (PA-040), PGN-007, SCY-635, SB-9200, SCB-719, TR-452,TEV-90110, TEV-90112, TEV-90111, TEV-90113, RN-18, Immuglo, and VIR-576.

Gene Therapy and Cell Therapy

Gene Therapy and Cell Therapy include the genetic modification tosilence a gene; genetic approaches to directly kill the infected cells;the infusion of immune cells designed to replace most of the subject'sown immune system to enhance the immune response to infected cells, oractivate the subject's own immune system to kill infected cells, or findand kill the infected cells; genetic approaches to modify cellularactivity to further alter endogenous immune responsiveness against theinfection.

Examples of dendritic cell therapy include AGS-004.

Gene Editors

Examples of gene editing systems include a CRISPR/Cas9 system, a zincfinger nuclease system, a TALEN system, a homing endonucleases system,and a meganuclease system.

Examples of HIV Targeting CRISPR/Cas9 Systems Include EBT101.

CAR-T Cell Therapy

CAR-T cell therapy includes a population of immune effector cellsengineered to express a chimeric antigen receptor (CAR), wherein the CARcomprises an HIV antigen-binding domain. The HIV antigen include an HIVenvelope protein or a portion thereof, gp120 or a portion thereof, a CD4binding site on gp120, the CD4-induced binding site on gp120, N glycanon gp120, the V2 of gp120, the membrane proximal region on gp41. Theimmune effector cell is a T cell or an NK cell. In some embodiments, theT cell is a CD4+ T cell, a CD8+ T cell, or a combination thereof.

Examples of HIV CAR-T include VC-CAR-T.

TCR-T Cell Therapy

TCR-T cell therapy includes T cells engineered to target HIV derivedpeptides present on the surface of virus-infected cells.

It will be appreciated by one of skill in the art that the additionaltherapeutic agents listed above may be included in more than one of theclasses listed above. The particular classes are not intended to limitthe functionality of those compounds listed in those classes.

In a specific embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with an HIVnucleoside or nucleotide inhibitor of reverse transcriptase and an HIVnon-nucleoside inhibitor of reverse transcriptase. In another specificembodiment, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with an HIV nucleoside ornucleotide inhibitor of reverse transcriptase, and an HIV proteaseinhibiting compound. In an additional embodiment, a compound disclosedherein, or a pharmaceutically acceptable salt thereof, is combined withan HIV nucleoside or nucleotide inhibitor of reverse transcriptase, anHIV non-nucleoside inhibitor of reverse transcriptase, and apharmacokinetic enhancer. In certain embodiments, a compound disclosedherein, or a pharmaceutically acceptable salt thereof, is combined withat least one HIV nucleoside inhibitor of reverse transcriptase, anintegrase inhibitor, and a pharmacokinetic enhancer. In anotherembodiment, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with two HIV nucleoside ornucleotide inhibitors of reverse transcriptase.

In a particular embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with one, two,three, four or more additional therapeutic agents selected from ATRIPLA®(efavirenz, tenofovir disoproxil fumarate, and emtricitabine); COMPLERA®(EVIPLERA®; rilpivirine, tenofovir disoproxil fumarate, andemtricitabine); STRIBILD® (elvitegravir, cobicistat, tenofovirdisoproxil fumarate, and emtricitabine); TRUVADA® (tenofovir disoproxilfumarate and emtricitabine; TDF+FTC); DESCOVY® (tenofovir alafenamideand emtricitabine); ODEFSEY® (tenofovir alafenamide, emtricitabine, andrilpivirine); GENVOYA® (tenofovir alafenamide, emtricitabine,cobicistat, and elvitegravir); BIKTARVY® (bictegravir, emtricitabine,tenofovir alafenamide); adefovir; adefovir dipivoxil; cobicistat;emtricitabine; tenofovir; tenofovir disoproxil; tenofovir disoproxilfumarate; tenofovir alafenamide; tenofovir alafenamide hemifumarate;TRIUMEQ® (dolutegravir, abacavir, and lamivudine); dolutegravir,abacavir sulfate, and lamivudine; raltegravir; raltegravir andlamivudine; maraviroc; enfuvirtide; ALUVIA® (KALETRA®; lopinavir andritonavir); COMBIVIR® (zidovudine and lamivudine; AZT+3TC); EPZICOM®(LIVEXA®; abacavir sulfate and lamivudine; ABC+3TC); TRIZIVIR® (abacavirsulfate, zidovudine, and lamivudine; ABC+AZT+3TC); rilpivirine;rilpivirine hydrochloride; atazanavir sulfate and cobicistat; atazanavirand cobicistat; darunavir and cobicistat; atazanavir; atazanavirsulfate; dolutegravir; elvitegravir; ritonavir; atazanavir sulfate andritonavir; darunavir; lamivudine; prolastin; fosamprenavir;fosamprenavir calcium efavirenz; etravirine; nelfinavir; nelfinavirmesylate; interferon; didanosine; stavudine; indinavir; indinavirsulfate; tenofovir and lamivudine; zidovudine; nevirapine; saquinavir;saquinavir mesylate; aldesleukin; zalcitabine; tipranavir; amprenavir;delavirdine; delavirdine mesylate; Radha-108 (receptol); lamivudine andtenofovir disoproxil fumarate; efavirenz, lamivudine, and tenofovirdisoproxil fumarate; phosphazid; lamivudine, nevirapine, and zidovudine;abacavir; and abacavir sulfate.

In a particular embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with abacavirsulfate, tenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate,tenofovir disoproxil hemifumarate, tenofovir alafenamide, tenofoviralafenamide hemifumarate, or bictegravir.

In a particular embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with tenofovir,tenofovir disoproxil, tenofovir disoproxil fumarate, tenofoviralafenamide, tenofovir alafenamide hemifumarate, or bictegravir.

In a particular embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with a firstadditional therapeutic agent selected from the group consisting ofabacavir sulfate, tenofovir, tenofovir disoproxil, tenofovir disoproxilfumarate, tenofovir alafenamide, tenofovir alafenamide hemifumarate, andbictegravir and a second additional therapeutic agent selected from thegroup consisting of emtricitabine and lamivudine.

In a particular embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with a firstadditional therapeutic agent selected from the group consisting oftenofovir, tenofovir disoproxil, tenofovir disoproxil fumarate,tenofovir alafenamide, tenofovir alafenamide hemifumarate, andbictegravir and a second additional therapeutic agent, wherein thesecond additional therapeutic agent is emtricitabine.

In certain embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with 5-30 mgtenofovir alafenamide fumarate, tenofovir alafenamide hemifumarate, ortenofovir alafenamide, and 200 mg emtricitabine. In certain embodiments,a compound disclosed herein, or a pharmaceutically acceptable saltthereof, is combined with 5-10, 5-15, 5-20, 5-25, 25-30, 20-30, 15-30,or 10-30 mg tenofovir alafenamide fumarate, tenofovir alafenamidehemifumarate, or tenofovir alafenamide, and 200 mg emtricitabine. Incertain embodiments, a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, is combined with 10 mg tenofovir alafenamidefumarate, tenofovir alafenamide hemifumarate, or tenofovir alafenamide,and 200 mg emtricitabine. In certain embodiments, a compound disclosedherein, or a pharmaceutically acceptable salt thereof, is combined with25 mg tenofovir alafenamide fumarate, tenofovir alafenamidehemifumarate, or tenofovir alafenamide, and 200 mg emtricitabine. Acompound as disclosed herein (e.g., a compound of Formula (I)) may becombined with the agents provided herein in any dosage amount of thecompound (e.g., from 1 mg to 500 mg of compound) the same as if eachcombination of dosages were specifically and individually listed.

In certain embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with 200-400 mgtenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, ortenofovir disoproxil, and 200 mg emtricitabine. In certain embodiments,a compound disclosed herein, or a pharmaceutically acceptable saltthereof, is combined with 200-250, 200-300, 200-350, 250-350, 250-400,350-400, 300-400, or 250-400 mg tenofovir disoproxil fumarate, tenofovirdisoproxil hemifumarate, or tenofovir disoproxil, and 200 mgemtricitabine. In certain embodiments, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with 300 mgtenofovir disoproxil fumarate, tenofovir disoproxil hemifumarate, ortenofovir disoproxil, and 200 mg emtricitabine. A compound as disclosedherein (e.g., a compound of Formula (I)) may be combined with the agentsprovided herein in any dosage amount of the compound (e.g., from 1 mg to500 mg of compound) the same as if each combination of dosages werespecifically and individually listed.

HBV Combination Therapy

In certain embodiments, a method for treating or preventing an HBVinfection in a human having or at risk of having the infection isprovided, comprising administering to the human a therapeuticallyeffective amount of a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, in combination with a therapeutically effectiveamount of one or more (e.g., one, two, three, four, one or two, one tothree, or one to four) additional therapeutic agents. In one embodiment,a method for treating an HBV infection in a human having or at risk ofhaving the infection is provided, comprising administering to the humana therapeutically effective amount of a compound disclosed herein, or apharmaceutically acceptable salt thereof, in combination with atherapeutically effective amount of one or more (e.g., one, two, three,four, one or two, one to three, or one to four) additional therapeuticagents.

In certain embodiments, the present disclosure provides a method fortreating an HBV infection, comprising administering to a patient in needthereof a therapeutically effective amount of a compound disclosedherein or a pharmaceutically acceptable salt thereof, in combinationwith a therapeutically effective amount of one or more (e.g., one, two,three, four, one or two, one to three, or one to four) additionaltherapeutic agents which are suitable for treating an HBV infection.

The compounds described herein may be used or combined with one or moreof a chemotherapeutic agent, an immunomodulator, an immunotherapeuticagent, a therapeutic antibody, a therapeutic vaccine, a bispecificantibody and “antibody-like” therapeutic protein (such as DARTs®,Duobodies®, Bites®, XmAbs®, TandAbs®, Fab derivatives), an antibody-drugconjugate (ADC), gene modifiers or gene editors (such as CRISPR Cas9,zinc finger nucleases, homing endonucleases, synthetic nucleases,TALENs), cell therapies such as CAR-T (chimeric antigen receptorT-cell), and TCR-T (an engineered T cell receptor) agent or anycombination thereof.

In certain embodiments, a compound of Formula (I) is formulated as atablet, which may optionally contain one or more other compounds usefulfor treating HBV. In certain embodiments, the tablet can contain anotheractive ingredient for treating HBV, such as 3-dioxygenase (IDO)inhibitors, Apolipoprotein A1 modulator, arginase inhibitors, B- andT-lymphocyte attenuator inhibitors, Bruton's tyrosine kinase (BTK)inhibitors, CCR2 chemokine antagonist, CD137 inhibitors, CD160inhibitors, CD305 inhibitors, CD4 agonist and modulator, compoundstargeting HBcAg, compounds targeting hepatitis B core antigen (HBcAg),core protein allosteric modulators, covalently closed circular DNA(cccDNA) inhibitors, cyclophilin inhibitors, cytotoxicT-lymphocyte-associated protein 4 (ipi4) inhibitors, DNA polymeraseinhibitor, Endonuclease modulator, epigenetic modifiers, Famesoid Xreceptor agonist, HBsAg inhibitors, HBsAg secretion or assemblyinhibitors, HBV DNA polymerase inhibitors, HBV replication inhibitors,HBV RNAse inhibitors, HBV viral entry inhibitors, HBx inhibitors,Hepatitis B large envelope protein modulator, Hepatitis B large envelopeprotein stimulator, Hepatitis B structural protein modulator, hepatitisB surface antigen (HBsAg) inhibitors, hepatitis B surface antigen(HBsAg) secretion or assembly inhibitors, hepatitis B virus E antigeninhibitors, hepatitis B virus replication inhibitors, Hepatitis virusstructural protein inhibitor, HIV-1 reverse transcriptase inhibitor,Hyaluronidase inhibitor, IAPs inhibitors, IL-2 agonist, IL-7 agonist,immunomodulators, indoleamine-2 inhibitors, inhibitors of ribonucleotidereductase, Interleukin-2 ligand, ipi4 inhibitors, lysine demethylaseinhibitors, histone demethylase inhibitors, KDM1 inhibitors, KDM5inhibitors, killer cell lectin-like receptor subfamily G member 1inhibitors, lymphocyte-activation gene 3 inhibitors, lymphotoxin betareceptor activators, modulators of Axl, modulators of B7-H3, modulatorsof B7-H4, modulators of CD160, modulators of CD161, modulators of CD27,modulators of CD47, modulators of CD70, modulators of GITR, modulatorsof HEVEM, modulators of ICOS, modulators of Mer, modulators of NKG2A,modulators of NKG2D, modulators of OX40, modulators of SIRPalpha,modulators of TIGIT, modulators of Tim-4, modulators of Tyro,Na+-taurocholate cotransporting polypeptide (NTCP) inhibitors, naturalkiller cell receptor 2B4 inhibitors, NOD2 gene stimulator, Nucleoproteininhibitor, nucleoprotein modulators, PD-1 inhibitors, PD-L1 inhibitors,Peptidylprolyl isomerase inhibitor, phosphatidylinositol-3 kinase (PI3K)inhibitors, Retinoic acid-inducible gene 1 stimulator, Reversetranscriptase inhibitor, Ribonuclease inhibitor, RNA DNA polymeraseinhibitor, SLC10A1 gene inhibitor, SMAC mimetics, Src tyrosine kinaseinhibitor, stimulator of interferon gene (STING) agonists, stimulatorsof NOD1, T cell surface glycoprotein CD28 inhibitor, T-cell surfaceglycoprotein CD8 modulator, Thymosin agonist, Thymosin alpha 1 ligand,Tim-3 inhibitors, TLR-3 agonist, TLR-7 agonist, TLR-9 agonist, TLR9 genestimulator, toll-like receptor (TLR) modulators, Viral ribonucleotidereductase inhibitor, and combinations thereof.

HBV Combination Drugs

Examples of combination drugs for the treatment of HBV include TRUVADA®(tenofovir disoproxil fumarate and emtricitabine); ABX-203, lamivudine,and PEG-IFN-alpha; ABX-203 adefovir, and PEG-IFNalpha; and INO-1800(INO-9112 and RG7944).

Other HBV Drugs

Examples of other drugs for the treatment of HBV includealpha-hydroxytropolones, amdoxovir, beta-hydroxycytosine nucleosides,AL-034, CCC-0975, elvucitabine, ezetimibe, cyclosporin A, gentiopicrin(gentiopicroside), JNJ-56136379, nitazoxanide, birinapant, NJK14047,NOV-205 (molixan, BAM-205), oligotide, mivotilate, feron, GST-HG-131,levamisole, Ka Shu Ning, alloferon, WS-007, Y-101 (Ti Fen Tai),rSIFN-co, PEG-IIFNm, KW-3, BP-Inter-014, oleanolic acid, HepB-nRNA,cTP-5 (rTP-5), HSK-II-2, HEISCO-106-1, HEISCO-106, Hepbarna, IBPB-006IA,Hepuyinfen, DasKloster 0014-01, ISA-204, Jiangantai (Ganxikang),MIV-210, OB-AI-004, PF-06, picroside, DasKloster-0039, hepulantai,IMB-2613, TCM-800B, reduced glutathione, RO-6864018, RG-7834, UB-551,and ZH-2N, and the compounds disclosed in US20150210682, (Roche), US2016/0122344 (Roche), WO2015173164, WO2016023877, US2015252057A (Roche),WO16128335A1 (Roche), WO16120186A1 (Roche), US2016237090A (Roche),WO16107833A1 (Roche), WO16107832A1 (Roche), US2016176899A (Roche),WO16102438A1 (Roche), WO16012470A1 (Roche), US2016220586A (Roche), andUS2015031687A (Roche).

HBV Vaccines

HBV vaccines include both prophylactic and therapeutic vaccines.Examples of HBV prophylactic vaccines include Vaxelis, Hexaxim,Heplisav, Mosquirix, DTwP-HBV vaccine, Bio-Hep-B, D/T/P/HBV/M(LBVP-0101; LBVW-0101), DTwP-Hepb-Hib-IPV vaccine, Heberpenta L,DTwP-HepB-Hib, V-419, CVI-HBV-001, Tetrabhay, hepatitis B prophylacticvaccine (Advax Super D), Hepatrol-07, GSK-223192A, ENGERIX B®,recombinant hepatitis B vaccine (intramuscular, Kangtai BiologicalProducts), recombinant hepatitis B vaccine (Hansenual polymorpha yeast,intramuscular, Hualan Biological Engineering), recombinant hepatitis Bsurface antigen vaccine, Bimmugen, Euforavac, Eutravac,anrix-DTaP-IPV-Hep B, HBAI-20, Infanrix-DTaP-IPV-Hep B-Hib, PentabioVaksin DTP-HB-Hib, Comvac 4, Twinrix, Euvax-B, Tritanrix HB, InfanrixHep B, Comvax, DTP-Hib-HBV vaccine, DTP-HBV vaccine, Yi Tai, HeberbiovacHB, Trivac HB, GerVax, DTwP-Hep B-Hib vaccine, Bilive, Hepavax-Gene,SUPERVAX, Comvac5, Shanvac-B, Hebsulin, Recombivax HB, Revac B mcf,Revac B+, Fendrix, DTwP-HepB-Hib, DNA-001, Shan5, Shan6, rhHBsAGvaccine, HBI pentavalent vaccine, LBVD, Infanrix HeXa, and DTaP-rHB-Hibvaccine.

Examples of HBV therapeutic vaccines include HBsAG-HBIG complex,ARB-1598, Bio-Hep-B, NASVAC, abi-HB (intravenous), ABX-203, Tetrabhay,GX-110E, GS-4774, peptide vaccine (epsilonPA-44), Hepatrol-07, NASVAC(NASTERAP), IMP-321, BEVAC, Revac B mcf, Revac B+, MGN-1333, KW-2,CVI-HBV-002, AltraHepB, VGX-6200, FP-02, FP-02.2, TG-1050, NU-500,HBVax, im/TriGrid/antigen vaccine, Mega-CD40L-adjuvanted vaccine,HepB-v, RG7944 (INO-1800), recombinant VLP-based therapeutic vaccine(HBV infection, VLP Biotech), AdTG-17909, AdTG-17910 AdTG-18202,ChronVac-B, TG-1050, and Lm HBV.

HBV DNA Polymerase Inhibitors

Examples of HBV DNA polymerase inhibitors include adefovir (HEPSERA®),emtricitabine (EMTRIVA®), tenofovir disoproxil fumarate (VIREAD®),tenofovir alafenamide, tenofovir, tenofovir disoproxil, tenofoviralafenamide fumarate, tenofovir alafenamide hemifumarate, tenofovirdipivoxil, tenofovir dipivoxil fumarate, tenofovir octadecyloxyethylester, CMX-157, besifovir, entecavir (BARACLUDE®), entecavir maleate,telbivudine (TYZEKA®), filocilovir, pradefovir, clevudine, ribavirin,lamivudine (EPIVIR-HBV®), phosphazide, famciclovir, fusolin, metacavir,SNC-019754, FMCA, AGX-1009, AR-11-04-26, HIP-1302, tenofovir disoproxilaspartate, tenofovir disoproxil orotate, and HS-10234.

Immunomodulators

Examples of immunomodulators include rintatolimod, imidol hydrochloride,ingaron, dermaVir, plaquenil (hydroxychloroquine), proleukin,hydroxyurea, mycophenolate mofetil (MPA) and its ester derivativemycophenolate mofetil (MMF), JNJ-440, WF-10, AB-452, ribavirin, IL-12,INO-9112, polymer polyethyleneimine (PEI), Gepon, VGV-1, MOR-22,CRV-431, JNJ-0535, TG-1050, ABI-H2158, BMS-936559, GS-9688, RO-7011785,RG-7854, AB-506, RO-6871765, AIC-649, and IR-103.

Toll-like Receptor (TLR) Modulators

TLR modulators include modulators of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6,TLR7, TLR8, TLR9, TLR10, TLR11, TLR12, and TLR13. Examples of TLR3modulators include rintatolimod, poly-ICLC, RIBOXXON®, Apoxxim,RIBOXXIM®, IPH-33, MCT-465, MCT-475, and ND-1.1.

Examples of TLR7 modulators include GS-9620, GSK-2245035, imiquimod,resiquimod, DSR-6434, DSP-3025, IMO-4200, MCT-465, MEDI-9197, 3M-051,SB-9922, 3M-052, Limtop, D, telratolimod, SP-0509, TMX-30X, TMX-202,RG-7863, RG-7795, LHC-165, RG-7854, and the compounds disclosed inUS20100143301 (Gilead Sciences), US20110098248 (Gilead Sciences), andUS20090047249 (Gilead Sciences).

Examples of TLR8 modulators include motolimod, resiquimod, 3M-051,3M-052, MCT-465, IMO-4200, VTX-763, VTX-1463, GS-9688 and the compoundsdisclosed in US20140045849 (Janssen), US20140073642 (Janssen),WO2014/056953 (Janssen), WO2014/076221 (Janssen), WO2014/128189(Janssen), US20140350031 (Janssen), WO2014/023813 (Janssen),US20080234251 (Array Biopharma), US20080306050 (Array Biopharma),US20100029585 (Ventirx Pharma), US20110092485 (Ventirx Pharma),US20110118235 (Ventirx Pharma), US20120082658 (Ventirx Pharma),US20120219615 (Ventirx Pharma), US20140066432 (Ventirx Pharma),US20140088085 (Ventirx Pharma), US20140275167 (Novira Therapeutics),US20130251673 (Novira Therapeutics), U.S. Pat. No. 9,670,205,US20160289229. U.S. patent application Ser. No. 15/692,161, and U.S.patent application Ser. No. 15/692,093.

Examples of TLR9 modulators include BB-001, BB-006, CYT-003, IMO-2055,IMO-2125, IMO-3100, IMO-8400, IR-103, IMO-9200, agatolimod, DIMS-9054,DV-1079, DV-1179, AZD-1419, leftolimod (MGN-1703), litenimod, andCYT-003-QbG10.

Examples of TLR7, TLR8 and TLR9 modulators include the compoundsdisclosed in WO2017047769 (Teika Seiyaku), WO2015014815 (Janssen),WO2018045150 (Gilead Sciences Inc), WO2018045144 (Gilead Sciences Inc),WO2015162075 (Roche), WO2017034986 (University of Kansas), WO2018095426(Jiangsu Hengrui Medicine Co Ltd), WO2016091698 (Roche), WO2016075661(GlaxoSmithKline Biologicals), WO2016180743 (Roche), WO2018089695(Dynavax Technologies), WO2016055553 (ROche), WO2015168279 (Novartis),WO2016107536 (Medshine Discovery), WO2018086593 (Livo (Shanghai)Pharmaceutical), WO2017106607 (Merck), WO2017061532 (Sumitomo DainipponPharma), WO2016023511 (Chia Tai Tianqing Pharmaceutical), WO2017076346(Chia Tai Tianqing Pharmaceutical), WO2017046112 (ROche), WO2018078149(Roche), WO2017040233 (3M Co), WO2016141092 (Gilead Sciences),WO2018049089 (BristolMyers Squibb), WO2015057655 (Eisai Co Ltd),WO2017001307 (Roche), WO2018005586 (BristolMyers Squibb), WO201704023(3M Co), WO2017163264 (Council of Scientific and Industrial Research(India)), WO2018046460 (GlaxoSmithKline Biologicals), WO2018047081(Novartis), WO2016142250 (Roche), WO2015168269 (Novartis), WO201804163(Roche), WO2018038877 (3M Co), WO2015057659 (Eisai Co Ltd), WO2017202704(Roche), WO2018026620 (BristolMyers Squibb), WO2016029077 (JanusBiotherapeutics), WO201803143 (Merck), WO2016096778 (Roche),WO2017190669 (Shanghai De Novo Pharmatech), U.S. Ser. No. 09/884,866(University of Minnesota), WO2017219931 (Sichuan KelunBiotechBiopharmaceutical), WO2018002319 (Janssen Sciences), WO2017216054(Roche), WO2017202703 (Roche), WO2017184735 (IFM Therapeutics),WO2017184746 (IFM Therapeutics), WO2015088045 (Takeda Pharmaceutical),WO2017038909 (Takeda Pharmaceutical), WO2015095780 (University ofKansas), WO2015023958 (University of Kansas)

Interferon Alpha Receptor Ligands

Examples of interferon alpha receptor ligands include interferonalpha-2b (INTRON A®), pegylated interferon alpha-2a (PEGASYS®),PEGylated interferon alpha-1b, interferon alpha 1b (HAPGEN®), Veldona,Infradure, Roferon-A, YPEG-interferon alfa-2a (YPEG-rhlFNalpha-2a),P-1101, Algeron, Alfarona, Ingaron (interferon gamma), rSIFN-co(recombinant super compound interferon), Ypeginterferon alfa-2b(YPEG-rhlFNalpha-2b), MOR-22, peginterferon alfa-2b (PEG-INTRON®),Bioferon, Novaferon, Inmutag (Inferon), MULTIFERON®, interferon alfa-nl(HUMOFERON®), interferon beta-1a (AVONEX®), Shaferon, interferon alfa-2b(Axxo), Alfaferone, interferon alfa-2b (BioGeneric Pharma),interferon-alpha 2 (CJ), Laferonum, VIPEG, BLAUFERON-A, BLAUFERON-B,Intermax Alpha, Realdiron, Lanstion, Pegaferon, PDferon-B PDferon-B,interferon alfa-2b (IFN, Laboratorios Bioprofarma), alfainterferona 2b,Kalferon, Pegnano, Feronsure, PegiHep, interferon alfa 2b(Zydus-Cadila), interferon alfa 2a, Optipeg A, Realfa 2B, Reliferon,interferon alfa-2b (Amega), interferon alfa-2b (Virchow),ropeginterferon alfa-2b, rHSA-IFN alpha-2a (recombinant human serumalbumin intereferon alpha 2a fusion protein), rHSA-IFN alpha 2b,recombinant human interferon alpha-(1b, 2a, 2b), peginterferon alfa-2b(Amega), peginterferon alfa-2a, Reaferon-EC, Proquiferon, Uniferon,Urifron, interferon alfa-2b (Changchun Institute of BiologicalProducts), Anterferon, Shanferon, Layfferon, Shang Sheng Lei Tai,INTEFEN, SINOGEN, Fukangtai, Pegstat, rHSA-IFN alpha-2b, SFR-9216, andInterapo (Interapa).

Hyaluronidase Inhibitors

Examples of hyaluronidase inhibitors include astodrimer.

Hepatitis B Surface Antigen (HBsAg) Inhibitors

Examples of HBsAg inhibitors include HBF-0259, PBHBV-001, PBHBV-2-15,PBHBV-2-1, REP-9AC, REP-9C, REP-9, REP-2139, REP-2139-Ca, REP-2165,REP-2055, REP-2163, REP-2165, REP-2053, REP-2031 and REP-006, andREP-9AC′.

Examples of HBsAg secretion inhibitors include BM601.

Cytotoxic T-Lymphocyte-Associated Protein 4 (Ipi4) Inhibitors

Examples of Cytotoxic T-lymphocyte-associated protein 4 (ipi4)inhibitors include AGEN-2041, AGEN-1884, ipilumimab, belatacept,PSI-001, PRS-010, Probody mAbs, tremelimumab, and JHL-1155.

Cyclophilin Inhibitors

Examples of cyclophilin inhibitors include CPI-431-32, EDP-494, OCB-030,SCY-635, NVP-015, NVP-018, NVP-019, STG-175, and the compounds disclosedin U.S. Pat. No. 8,513,184 (Gilead Sciences), US20140030221 (GileadSciences), US20130344030 (Gilead Sciences), and US20130344029 (GileadSciences).

HBV Viral Entry Inhibitors

Examples of HBV viral entry inhibitors include Myrcludex B.

Antisense Oligonucleotide Targeting Viral mRNA

Examples of antisense oligonucleotide targeting viral mRNA includeISIS-HBVRx, IONIS-HBVRx, IONIS-GSK6-LRx, GSK-3389404, RG-6004.

Short Interfering RNAs (siRNA) and ddRNAi.

Examples of siRNA include TKM-HBV (TKM-HepB), ALN-HBV, SR-008,HepB-nRNA, and ARC-520, ARC-521, ARB-1740, ARB-1467.

Examples of DNA-directed RNA interference (ddRNAi) include BB-HB-331.

Endonuclease Modulators

Examples of endonuclease modulators include PGN-514.

Ribonucelotide Reductase Inhibitors

Examples of inhibitors of ribonucleotide reductase include Trimidox.

HBV E Antigen Inhibitors

Examples of HBV E antigen inhibitors include wogonin.

Covalently Closed Circular DNA (cccDNA) Inhibitors

Examples of cccDNA inhibitors include BSBI-25, and CHR-101.

Farnesoid X receptor agonist

Examples of famesoid x receptor agonist such as EYP-001, GS-9674,EDP-305, MET-409, Tropifexor, AKN-083, RDX-023, BWD-100, LMB-763, INV-3,NTX-023-1, EP-024297 and GS-8670

HBV Antibodies

Examples of HBV antibodies targeting the surface antigens of thehepatitis B virus include GC-1102, XTL-17, XTL-19, KN-003, IV HepabulinSN, and fully human monoclonal antibody therapy (hepatitis B virusinfection, Humabs BioMed).

Examples of HBV antibodies, including monoclonal antibodies andpolyclonal antibodies, include Zutectra, Shang Sheng Gan Di, Uman Big(Hepatitis B Hyperimmune), Omri-Hep-B, Nabi-HB, Hepatect CP, HepaGam B,igantibe, Niuliva, CT-P24, hepatitis B immunoglobulin (intravenous, pH4,HBV infection, Shanghai RAAS Blood Products), and Fovepta (BT-088).

Fully human monoclonal antibodies include HBC-34.

CCR2 Chemokine Antagonists

Examples of CCR2 chemokine antagonists include propagermanium.

Thymosin Agonists

Examples of thymosin agonists include Thymalfasin, recombinant thymosinalpha 1 (GeneScience)

Cytokines

Examples of cytokines include recombinant IL-7, CYT-107, interleukin-2(IL-2, Immunex), recombinant human interleukin-2 (Shenzhen Neptunus),IL-15, IL-21, IL-24, and celmoleukin.

Nucleoprotein Modulators

Nucleoprotein modulators may be either HBV core or capsid proteininhibitors. Examples of nucleoprotein modulators include GS-4882,AB-423, AT-130, GLS4, NVR-1221, NVR-3778, AL-3778, BAY 41-4109,morphothiadine mesilate, ARB-168786, ARB-880, JNJ-379, RG-7907,HEC-72702, AB-506, ABI-H0731, JNJ-440, ABI-H2158 and DVR-23.

Examples of capsid inhibitors include the compounds disclosed inUS20140275167 (Novira Therapeutics), US20130251673 (NoviraTherapeutics), US20140343032 (Roche), WO2014037480 (Roche),US20130267517 (Roche), WO2014131847 (Janssen), WO2014033176 (Janssen),WO2014033170 (Janssen), WO2014033167 (Janssen), WO2015/059212 (Janssen),WO2015118057 (Janssen), WO2015011281 (Janssen), WO2014184365 (Janssen),WO2014184350 (Janssen), WO2014161888 (Janssen), WO2013096744 (Novira),US20150225355 (Novira), US20140178337 (Novira), US20150315159 (Novira),US20150197533 (Novira), US20150274652 (Novira), US20150259324, (Novira),US20150132258 (Novira), U.S. Pat. No. 9,181,288 (Novira), WO2014184350(Janssen), WO2013144129 (Roche), WO2017198744 (Roche), US 20170334882(Novira), US 20170334898 (Roche), WO2017202798 (Roche), WO2017214395(Enanta), WO2018001944 (Roche), WO2018001952 (Roche), WO2018005881(Novira), WO2018005883 (Novira), WO2018011100 (Roche), WO2018011160(Roche), WO2018011162 (Roche), WO2018011163 (Roche), WO2018036941(Roche), WO2018043747 (Kyoto Univ), US20180065929 (Janssen),WO2016168619 (Indiana University), WO2016195982 (The Penn StateFoundation), WO2017001655 (Janssen), WO2017048950 (AssemblyBiosciences), WO2017048954 (Assembly Biosciences), WO2017048962(Assembly Biosciences), US20170121328 (Novira), US20170121329 (Novira).

Examples of transcript inhibitors include the compounds disclosed inWO2017013046 (Roche), WO2017016960 (Roche), WO2017017042 (Roche),WO2017017043 (Roche), WO2017061466 (Toyoma chemicals), WO2016177655(Roche), WO2016161268 (Enanta). WO2017001853 (Redex Pharma),WO2017211791 (Roche), WO2017216685 (Novartis), WO2017216686 (Novartis),WO2018019297 (Ginkgo Pharma), WO2018022282 (Newave Pharma),US20180030053 (Novartis), WO2018045911 (Zhejiang Pharma).

Retinoic Acid-Inducible Gene 1 Stimulators

Examples of stimulators of retinoic acid-inducible gene 1 includeSB-9200, SB-40, SB-44, ORI-7246, ORI-9350, ORI-7537, ORI-9020, ORI-9198,and ORI-7170, RGT-100.

NOD2 Stimulators

Examples of stimulators of NOD2 include SB-9200.

Phosphatidylinositol 3-Kinase (PI3K) Inhibitors

Examples of PI3K inhibitors include idelalisib, ACP-319, AZD-8186,AZD-8835, buparlisib, CDZ-173, CLR-457, pictilisib, neratinib,rigosertib, rigosertib sodium, EN-3342, TGR-1202, alpelisib, duvelisib,IPI-549, UCB-5857, taselisib, XL-765, gedatolisib, ME-401, VS-5584,copanlisib, CAI orotate, perifosine, RG-7666, GSK-2636771, DS-7423,panulisib, GSK-2269557, GSK-2126458, CUDC-907, PQR-309, INCB-40093,pilaralisib, BAY-1082439, puquitinib mesylate, SAR-245409, AMG-319,RP-6530, ZSTK-474, MLN-1117, SF-1126, RV-1729, sonolisib, LY-3023414,SAR-260301, TAK-117, HMPL-689, tenalisib, voxtalisib, and CLR-1401.

Indoleamine-2, 3-dioxygenase (IDO) Pathway Inhibitors

Examples of IDO inhibitors include epacadostat (INCB24360), resminostat(4SC-201), indoximod, F-001287, SN-35837, NLG-919, GDC-0919, GBV-1028,GBV-1012, NKTR-218, and the compounds disclosed in US20100015178(Incyte), US2016137652 (Flexus Biosciences, Inc.), WO2014073738 (FlexusBiosciences, Inc.), and WO2015188085 (Flexus Biosciences, Inc.).

PD-1 Inhibitors

Examples of PD-1 inhibitors include cemiplimab, nivolumab,pembrolizumab, pidilizumab, BGB-108, STI-A1014, SHR-1210, PDR-001,PF-06801591, IBI-308, GB-226, STI-1110, JNJ-63723283, CA-170,durvalumab, atezolizumab and mDX-400, JS-001, Camrelizumab, Sintilimab,Sintilimab, tislelizumab, BCD-100, BGB-A333 JNJ-63723283, GLS-010(WBP-3055), CX-072, AGEN-2034, GNS-1480 (Epidermal growth factorreceptor antagonist; Programmed cell death ligand 1 inhibitor), CS-1001,M-7824 (PD-L1/TGF-β bifunctional fusion protein), Genolimzumab,BMS-936559

PD-L1 Inhibitors

Examples of PD-L1 inhibitors include atezolizumab, avelumab, AMP-224,MEDI-0680, RG-7446, GX-P2, durvalumab, KY-1003, KD-033, MSB-0010718C,TSR-042, ALN-PDL, STI-A1014, GS-4224, CX-072, and BMS-936559.

Examples of PD-1 inhibitors include the compounds disclosed inWO2017112730 (Incyte Corp), WO2017087777 (Incyte Corp), WO2017017624,WO2014151634 (BristolMyers Squibb Co), WO201317322 (BristolMyers SquibbCo), WO2018119286 (Incyte Corp), WO2018119266 (Incyte Corp),WO2018119263 (Incyte Corp), WO2018119236 (Incyte Corp), WO2018119221(Incyte Corp), WO2018118848 (BristolMyers Squibb Co), WO20161266460(BristolMyers Squibb Co), WO2017087678 (BristolMyers Squibb Co),WO2016149351 (BristolMyers Squibb Co), WO2015033299 (Aurigene DiscoveryTechnologies Ltd), WO2015179615 (Eisai Co Ltd; Eisai ResearchInstitute), WO2017066227 (BristolMyers Squibb Co), WO2016142886(Aurigene Discovery Technologies Ltd), WO2016142852 (Aurigene DiscoveryTechnologies Ltd), WO2016142835 (Aurigene Discovery Technologies Ltd;Individual), WO2016142833 (Aurigene Discovery Technologies Ltd),WO2018085750 (BristolMyers Squibb Co), WO2015033303 (Aurigene DiscoveryTechnologies Ltd), WO2017205464 (Incyte Corp), WO2016019232 (3M Co;Individual; Texas A&M University System), WO2015160641 (BristolMyersSquibb Co), WO2017079669 (Incyte Corp), WO2015033301 (Aurigene DiscoveryTechnologies Ltd), WO2015034820 (BristolMyers Squibb Co), WO2018073754(Aurigene Discovery Technologies Ltd), WO2016077518 (BristolMyers SquibbCo), WO2016057624 (BristolMyers Squibb Co), WO2018044783 (Incyte Corp),WO2016100608 (BristolMyers Squibb Co), WO2016100285 (BristolMyers SquibbCo), WO2016039749 (BristolMyers Squibb Co), WO2015019284 (CambridgeEnterprise Ltd), WO2016142894 (Aurigene Discovery Technologies Ltd),WO2015134605 (BristolMyers Squibb Co), WO2018051255 (Aurigene DiscoveryTechnologies Ltd), WO2018051254 (Aurigene Discovery Technologies Ltd),WO2017222976 (Incyte Corp), WO2017070089 (Incyte Corp), WO2018044963(BristolMyers Squibb Co), WO2013144704 (Aurigene Discovery TechnologiesLtd), WO2018013789 (Incyte Corp), WO2017176608 (BristolMyers Squibb Co),WO2018009505 (BristolMyers Squibb Co), WO2011161699 (Aurigene DiscoveryTechnologies Ltd), WO2015119944 (Incyte Corp; Merck Sharp & Dohme Corp),WO2017192961 (Incyte Corp), WO2017106634 (Incyte Corp), WO2013132317(Aurigene Discovery Technologies Ltd), WO2012168944 (Aurigene DiscoveryTechnologies Ltd), WO2015036927 (Aurigene Discovery Technologies Ltd),WO2015044900 (Aurigene Discovery Technologies Ltd), WO2018026971(Arising International).

Recombinant Thymosin Alpha-1

Examples of recombinant thymosin alpha-1 include NL-004 and PEGylatedthymosin alpha-1.

Bruton's Tyrosine Kinase (BTK) Inhibitors

Examples of BTK inhibitors include ABBV-105, acalabrutinib (ACP-196),ARQ-531, BMS-986142, dasatinib, ibrutinib, GDC-0853, PRN-1008, SNS-062,ONO-4059, BGB-3111, ML-319, MSC-2364447, RDX-022, X-022, AC-058,RG-7845, spebrutinib, TAS-5315, TP-0158, TP-4207, HM-71224, KBP-7536,M-2951, TAK-020, AC-0025, and the compounds disclosed in US20140330015(Ono Pharmaceutical), US20130079327 (Ono Pharmaceutical), andUS20130217880 (Ono Pharmaceutical).

KDM Inhibitors

Examples of KDM5 inhibitors include the compounds disclosed inWO2016057924 (Genentech/Constellation Pharmaceuticals), US20140275092(Genentech/Constellation Pharmaceuticals), US20140371195(Epitherapeutics) and US20140371214 (Epitherapeutics), US20160102096(Epitherapeutics), US20140194469 (Quanticel), US20140171432,US20140213591 (Quanticel), US20160039808 (Quanticel), US20140275084(Quanticel), WO2014164708 (Quanticel).

Examples of KDM1 inhibitors include the compounds disclosed in U.S. Pat.No. 9,186,337B2 (Oryzon Genomics), GSK-2879552, and RG-6016.

STING Agonists

Examples of STING agonists include SB-11285, AdVCA0848, STINGVAX, amdthe compounds disclosed in WO 2018065360 (“Biolog Life Science InstituteForschungslabor und Biochemica-Vertrieb GmbH, Germany), WO 2018009466(Aduro Biotech), WO 2017186711 (InvivoGen), WO 2017161349 (ImmuneSensor), WO 2017106740 (Aduro Biotech), US 20170158724 (GlaxoSmithkiline), WO 2017075477 (Aduro Biotech), US 20170044206 (Merck), WO2014179760 (University of California), WO2018098203 (Janssn),WO2018118665 (Merck), WO2018118664 (Merck), WO2018100558 (Takeda),WO2018067423 (Merck), WO2018060323 (Boehringer).

Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTI)

Examples of NNRTI include the compounds disclosed in WO2018118826(Merck), WO2018080903 (Merck), WO2018119013 (Merck), WO2017100108(Idenix), WO2017027434 (Merck), WO2017007701 (Merck), WO2008005555(Gilead).

HBV Replication Inhibitors

Examples of hepatitis B virus replication inhibitors includeisothiafludine, IQP-HBV, RM-5038, and Xingantie.

Arginase Inhibitors

Examples of Arginase inhibitors include CB-1158, C-201, and resminostat.

Gene Therapy and Cell Therapy

Gene therapy and cell therapy includes the genetic modification tosilence a gene; genetic approaches to directly kill the infected cells;the infusion of immune cells designed to replace most of the patient'sown immune system to enhance the immune response to infected cells, oractivate the patient's own immune system to kill infected cells, or findand kill the infected cells; and genetic approaches to modify cellularactivity to further alter endogenous immune responsiveness against theinfection.

Gene Editors

Examples of genome editing systems include a CRISPR/Cas9 system, a zincfinger nuclease system, a TALEN system, a homing endonucleases system,and a meganuclease system; e.g., cccDNA elimination via targetedcleavage, and altering one or more of the hepatitis B virus (HBV) viralgenes. Altering (e.g., knocking out and/or knocking down) the PreC, C,X, PreSI, PreS2, S, P or SP gene refers to (1) reducing or eliminatingPreC, C, X, PreSI, PreS2, S, P or SP gene expression, (2) interferingwith Precore, Core, X protein, Long surface protein, middle surfaceprotein, S protein (also known as HBs antigen and HBsAg), polymeraseprotein, and/or Hepatitis B spliced protein function (HBe, HBc, HBx,PreS1, PreS2, S, Pol, and/or HBSP or (3) reducing or eliminating theintracellular, serum and/or intraparenchymal levels of HBe, HBc, HBx,LHBs, MHBs, SHBs, Pol, and/or HBSP proteins. Knockdown of one or more ofthe PreC, C, X, PreSI, PreS2, S, P and/or SP gene(s) is performed bytargeting the gene(s) within HBV cccDNA and/or integrated HBV DNA.

CAR-T Cell Therapy

CAR T cell therapy includes a population of immune effector cellsengineered to express a chimeric antigen receptor (CAR), wherein the CARcomprises an HBV antigen-binding domain. The immune effector cell is a Tcell or an NK cell. In some embodiments, the T cell is a CD4+ T cell, aCD8+ T cell, or a combination thereof. Cells can be autologous orallogeneic.

TCR-T Cell Therapy

TCR T cell therapy includes T cells expressing HBV-specific T cellreceptors. TCR-T cells are engineered to target HBV derived peptidespresented on the surface of virus-infected cells. In some embodiments,the T-cells express HBV surface antigen (HBsAg)-specific TCR. Examplesof TCR-T therapy directed to treatment of HBV include LTCR-H2-1.

In another specific embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with an HBV DNApolymerase inhibitor, one or two additional therapeutic agents selectedfrom the group consisting of immunomodulators, TLR modulators, HBsAginhibitors, HBsAg secretion or assembly inhibitors, HBV therapeuticvaccines, HBV antibodies including HBV antibodies targeting the surfaceantigens of the hepatitis B virus and bispecific antibodies and“antibody-like” therapeutic proteins (such as DARTs®, DUOBODIES®,BITES®, XmAbs®, TandAbs®, Fab derivatives, or TCR-like antibodies),cyclophilin inhibitors, stimulators of retinoic acid-inducible gene 1,stimulators of RIG-I like receptors, PD-1 inhibitors, PD-L1 inhibitors,Arginase inhibitors, PI3K inhibitors, IDO inhibitors, and stimulators ofNOD2, and one or two additional therapeutic agents selected from thegroup consisting of HBV viral entry inhibitors, NTCP inhibitors, HBxinhibitors, cccDNA inhibitors, HBV antibodies targeting the surfaceantigens of the hepatitis B virus, siRNA, miRNA gene therapy agents,sshRNAs, KDM5 inhibitors, and nucleoprotein modulators (HBV core orcapsid protein modulators).

In another specific embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with an HBV DNApolymerase inhibitor and at least a second additional therapeutic agentselected from the group consisting of: immunomodulators, TLR modulators,HBsAg inhibitors, HBV therapeutic vaccines, HBV antibodies including HBVantibodies targeting the surface antigens of the hepatitis B virus andbispecific antibodies and “antibody-like” therapeutic proteins (such asDARTs®, DUOBODIES®, BITES®, XmAbs®, TandAbs®, Fab derivatives, orTCR-like antibodies), cyclophilin inhibitors, stimulators of retinoicacid-inducible gene 1, stimulators of RIG-I like receptors, PD-1inhibitors, PD-L1 inhibitors, Arginase inhibitors, PI3K inhibitors, IDOinhibitors, and stimulators of NOD2.

In another specific embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with an HBV DNApolymerase inhibitor and at least a second additional therapeutic agentselected from the group consisting of: HBV viral entry inhibitors, NTCPinhibitors, HBx inhibitors, cccDNA inhibitors, HBV antibodies targetingthe surface antigens of the hepatitis B virus, siRNA, miRNA gene therapyagents, sshRNAs, KDM5 inhibitors, and nucleoprotein modulators (HBV coreor capsid protein inhibitors).

In a particular embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is combined with compoundssuch as those disclosed in U.S. Publication No. 2010/0143301 (GileadSciences), U.S. Publication No. 2011/0098248 (Gilead Sciences), U.S.Publication No. 2009/0047249 (Gilead Sciences), U.S. Pat. No. 8,722,054(Gilead Sciences), U.S. Publication No. 2014/0045849 (Janssen), U.S.Publication No. 2014/0073642 (Janssen), WO2014/056953 (Janssen),WO2014/076221 (Janssen), WO2014/128189 (Janssen), U.S. Publication No.2014/0350031 (Janssen), WO2014/023813 (Janssen), U.S. Publication No.2008/0234251 (Array Biopharma), U.S. Publication No. 2008/0306050 (ArrayBiopharma), U.S. Publication No. 2010/0029585 (Ventirx Pharma), U.S.Publication No. 2011/0092485 (Ventirx Pharma), US2011/0118235 (VentirxPharma), U.S. Publication No. 2012/0082658 (Ventirx Pharma), U.S.Publication No. 2012/0219615 (Ventirx Pharma), U.S. Publication No.2014/0066432 (Ventirx Pharma), U.S. Publication No. 2014/0088085(Ventirx Pharma), U.S. Publication No. 2014/0275167 (NoviraTherapeutics), U.S. Publication No. 2013/0251673 (Novira Therapeutics),U.S. Pat. No. 8,513,184 (Gilead Sciences), U.S. Publication No.2014/0030221 (Gilead Sciences), U.S. Publication No. 2013/0344030(Gilead Sciences), U.S. Publication No. 2013/0344029 (Gilead Sciences),US20140275167 (Novira Therapeutics), US20130251673 (NoviraTherapeutics), U.S. Publication No. 2014/0343032 (Roche), WO2014037480(Roche), U.S. Publication No. 2013/0267517 (Roche), WO2014131847(Janssen), WO2014033176 (Janssen), WO2014033170 (Janssen), WO2014033167(Janssen), WO2015/059212 (Janssen), WO2015118057 (Janssen), WO2015011281(Janssen), WO2014184365 (Janssen), WO2014184350 (Janssen), WO2014161888(Janssen), WO2013096744 (Novira), US20150225355 (Novira), US20140178337(Novira), US20150315159 (Novira), US20150197533 (Novira), US20150274652(Novira), US20150259324, (Novira), US20150132258 (Novira), U.S. Pat. No.9,181,288 (Novira), WO2014184350 (Janssen), WO2013144129 (Roche),US20100015178 (Incyte), US2016137652 (Flexus Biosciences, Inc.),WO2014073738 (Flexus Biosciences, Inc.), WO2015188085 (FlexusBiosciences, Inc.), U.S. Publication No. 2014/0330015 (OnoPharmaceutical), U.S. Publication No. 2013/0079327 (Ono Pharmaceutical),U.S. Publication No. 2013/0217880 (Ono pharmaceutical), WO2016057924(Genentech/Constellation Pharmaceuticals), US20140275092(Genentech/Constellation Pharmaceuticals), US20140371195(Epitherapeutics) and US20140371214 (Epitherapeutics), US20160102096(Epitherapeutics), US20140194469 (Quanticel), US20140171432,US20140213591 (Quanticel), US20160039808 (Quanticel), US20140275084(Quanticel), WO2014164708 (Quanticel), U.S. Pat. No. 9,186,337B2 (OryzonGenomics), and other drugs for treating HBV, and combinations thereof.

Cancer Combination Therapy

In one embodiment, the compound of the disclosure may be employed withother therapeutic methods of cancer treatment. Preferably, combinationtherapy with chemotherapeutic, hormonal, antibody, surgical and/orradiation treatments are contemplated.

In some embodiments, the further anti-cancer therapy is surgery and/orradiotherapy.

In some embodiments, the further anti-cancer therapy is at least oneadditional cancer medicament.

In some embodiments, there is provided a combination comprising acompound of Formula (, or a pharmaceutically acceptable salt thereof andat least one further cancer medicament.

In some embodiments, there is provided a combination comprising acompound of the present disclosure or a pharmaceutically acceptable saltthereof and at least one further cancer medicament, for use in therapy.

In some embodiments, there is provided the use of a combinationcomprising a compound of the present disclosure, or a pharmaceuticallyacceptable salt thereof and at least one cancer medicament, in themanufacture of a medicament for the treatment of cancer.

Examples of further cancer medicaments include intercalating substancessuch as anthracycline, doxorubicin, idarubicin, epirubicin, anddaunorubicin; topoisomerase inhibitors such as irinotecan, topotecan,camptothecin, lamellarin D, etoposide, teniposide, mitoxantrone,amsacrine, ellipticines and aurintricarboxylic acid; nitrosoureacompounds such as carmustine (BCNU), lomustine (CCNU), and streptozocin;nitrogen mustards such as cyclophosphamide, mechlorethamine, uramustine,bendamustine, melphalan, chlorambucil, mafosfamide, trofosfamid andifosfamide; alkyl sulfonates such as busulfan and treosulfan; alkylatingagents such as procarbazin, dacarbazin, temozolomid and thiotepa;platinum analogues such as cisplatin, carboplatin, nedaplatin,oxaliplatin, satraplatin, and triplatin tetranitrate; microtubuledisruptive drugs such as vinblastine, colcemid and nocodazole;antifolates like methotrexate, aminopterin, dichloromethotrexat,pemetrexed, raltitrexed and pralatrexate: purine analogues likeazathioprine, mercaptopurine, thioguanine, fludarabine, fludarabinephosphate, pentostatin and cladribine; pyrimidine analogues like5-fluorouracil, floxuridine, cytarabine, 6-azauracil, gemcitabine;steroids such as gestagene, androgene, glucocorticoids, dexamethasone,prednisolone, and prednisone; anti-cancer antibodies such as monoclonalantibodies, e.g., alemtuzumab, apolizumab, cetuximab, epratuzumab,galiximab, gemtuzumab, ipilimumab, labetuzumab, panitumumab, rituximab,trastuzumab, nimotuzumab, mapatumumab, matuzumab, rhMab ICR62 andpertuzumab, radioactively labeled antibodies and antibody-drugconjugates; anti-cancer peptides such as radioactively labeled peptidesand peptide-drug conjugates; and taxane and taxane analogues such aspaclitaxel and docetaxel.

In certain embodiments, a method for treating or preventing ahyperproliferative disorder or cancer in a human or animal having or atrisk of having the hyperproliferative disorder or cancer is provided,comprising administering to the human or animal a therapeuticallyeffective amount of a compound of the present disclosure as disclosedherein, or a pharmaceutically acceptable salt thereof, in combinationwith a therapeutically effective amount of one or more (e.g., one, two,three, one or two, or one to three) additional therapeutic agents. Inone embodiment, a method for treating a hyperproliferative disorder orcancer in a human or animal having or at risk of having thehyperproliferative disorder or cancer is provided, comprisingadministering to the human or animal a therapeutically effective amountof a compound disclosed herein, or a pharmaceutically acceptable saltthereof, in combination with a therapeutically effective amount of oneor more (e.g., one, two, three, one or two, or one to three) additionaltherapeutic agents.

In certain embodiments, the present disclosure provides a method fortreating a hyperproliferative disorder or cancer, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of a compound disclosed herein, or a pharmaceutically acceptablesalt thereof, in combination with a therapeutically effective amount ofone or more additional therapeutic agents which are suitable fortreating hyperproliferative disorder or cancer.

The compounds described herein may be used or combined with one or moreof a chemotherapeutic agent, an anti-cancer agent, an anti-angiogenicagent, an anti-fibrotic agent, an immunotherapeutic agent, a therapeuticantibody, a bispecific antibody and “antibody-like” therapeutic protein(such as DARTs®, Duobodies®, Bites®, XmAbs®, TandAbs®, Fab derivatives),an antibody-drug conjugate (ADC), a radiotherapeutic agent, ananti-neoplastic agent, an anti-proliferation agent, an oncolytic virus,a gene modifier or editor (such as CRISPR/Cas9, zinc finger nucleases orsynthetic nucleases, TALENs), a CAR (chimeric antigen receptor) T-cellimmunotherapeutic agent, an engineered T cell receptor (TCR-T), or anycombination thereof. These therapeutic agents may be in the forms ofcompounds, antibodies, polypeptides, or polynucleotides. In oneembodiment, provided herein is a product comprising a compound describedherein and an additional therapeutic agent as a combined preparation forsimultaneous, separate, or sequential use in therapy.

The one or more therapeutic agents include, but are not limited to, aninhibitor, agonist, antagonist, ligand, modulator, stimulator, blocker,activator or suppressor of a gene, ligand, receptor, protein, or factor.Non-limiting examples of additional therapeutic agents include: Abelsonmurine leukemia viral oncogene homolog 1 gene (ABL, such as ABL1),Acetyl-CoA carboxylase (such as ACC1/2), activated CDC kinase (ACK, suchas ACK1), Adenosine deaminase, adenosine receptor (such as A2B, A2a,A3), Adenylate cyclase, ADP ribosyl cyclase-1, adrenocorticotropichormone receptor (ACTH), Aerolysin, AKT1 gene, Alk-5 protein kinase,Alkaline phosphatase, Alpha 1 adrenoceptor, Alpha 2 adrenoceptor,Alpha-ketoglutarate dehydrogenase (KGDH), Aminopeptidase N, AMPactivated protein kinase, anaplastic lymphoma kinase (ALK, such asALK1), Androgen receptor, Angiopoietin (such as ligand-1, ligand-2),Angiotensinogen (AGT) gene, murine thymoma viral oncogene homolog 1(AKT) protein kinase (such as AKT1, AKT2, AKT3), apolipoprotein A-I(APOAl) gene, Apoptosis inducing factor, apoptosis protein (such as 1,2), apoptosis signal-regulating kinase (ASK, such as ASK1), Arginase(I), Arginine deiminase, Aromatase, Asteroid homolog 1 (ASTE1) gene,ataxia telangiectasia and Rad 3 related (ATR) serine/threonine proteinkinase, Aurora protein kinase (such as 1, 2), Axl tyrosine kinasereceptor, Baculoviral IAP repeat containing 5 (BIRC5) gene, Basigin,B-cell lymphoma 2 (BCL2) gene, Bcl2 binding component 3, Bcl2 protein,BCL2L11 gene, BCR (breakpoint cluster region) protein and gene, Betaadrenoceptor, Beta-catenin, B-lymphocyte antigen CD19, B-lymphocyteantigen CD20, B-lymphocyte cell adhesion molecule, B-lymphocytestimulator ligand, Bone morphogenetic protein-10 ligand, Bonemorphogenetic protein-9 ligand modulator, Brachyury protein, Bradykininreceptor, B-Raf proto-oncogene (BRAF), Brc-Abl tyrosine kinase,Bromodomain and external domain (BET) bromodomain containing protein(such as BRD2, BRD3, BRD4), Bruton's tyrosine kinase (BTK), Calmodulin,calmodulin-dependent protein kinase (CaMK, such as CAMKII), Cancertestis antigen 2, Cancer testis antigen NY-ESO-1, cancer/testis antigen1B (CTAG1) gene, Cannabinoid receptor (such as CB1, CB2), Carbonicanhydrase, casein kinase (CK, such as CKI, CKII), Caspase (such ascaspase-3, caspase-7, Caspase-9), caspase 8 apoptosis-related cysteinepeptidase CASP8-FADD-like regulator, Caspase recruitment domainprotein-15, Cathepsin G, CCR5 gene, CDK-activating kinase (CAK),Checkpoint kinase (such as CHK1, CHK2), chemokine (C-C motif) receptor(such as CCR2, CCR4, CCR5), chemokine (C—X-C motif) receptor (such asCXCR4, CXCR1 and CXCR2), Chemokine CC21 ligand, Cholecystokinin CCK2receptor, Chorionic gonadotropin, c-Kit (tyrosine-protein kinase Kit orCD117), Claudin (such as 6, 18), cluster of differentiation (CD) such asCD4, CD27, CD29, CD30, CD33, CD37, CD40, CD40 ligand receptor, CD40ligand, CD40LG gene, CD44, CD45, CD47, CD49b, CD51, CD52, CD55, CD58,CD66e, CD70 gene, CD74, CD79, CD79b, CD79B gene, CD80, CD95, CD99,CD117, CD122, CDw123, CD134, CDw137, CD158a, CD158b1, CD158b2, CD223,CD276 antigen; clusterin (CLU) gene, Clusterin, c-Met (hepatocyte growthfactor receptor (HGFR)), Complement C3, Connective tissue growth factor,COP9 signalosome subunit 5, CSF-1 (colony-stimulating factor 1receptor), CSF2 gene, CTLA-4 (cytotoxic T-lymphocyte protein 4)receptor, Cyclin Dl, Cyclin G1, cyclin-dependent kinases (CDK, such asCDK1, CDK1B, CDK2-9), cyclooxygenase (such as 1, 2), CYP2B1 gene,Cysteine palmitoyltransferase porcupine, Cytochrome P450 11B2,Cytochrome P450 17, cytochrome P450 17A1, Cytochrome P450 2D6,cytochrome P450 3A4, Cytochrome P450 reductase, cytokine signalling-1,cytokine signalling-3, Cytoplasmic isocitrate dehydrogenase, Cytosinedeaminase, cytosine DNA methyltransferase, cytotoxic T-lymphocyteprotein-4, DDR2 gene, Delta-like protein ligand (such as 3, 4),Deoxyribonuclease, Dickkopf-1 ligand, dihydrofolate reductase (DHFR),Dihydropyrimidine dehydrogenase, Dipeptidyl peptidase IV, discoidindomain receptor (DDR, such as DDR1), DNA binding protein (such asHU-beta), DNA dependent protein kinase, DNA gyrase, DNAmethyltransferase, DNA polymerase (such as alpha), DNA primase, dUTPpyrophosphatase, L-dopachrome tautomerase, echinoderm microtubule likeprotein 4, EGFR tyrosine kinase receptor, Elastase, Elongation factor 1alpha 2, Elongation factor 2, Endoglin, Endonuclease, Endoplasmin,Endosialin, Endostatin, endothelin (such as ET-A, ET-B), Enhancer ofzeste homolog 2 (EZH2), Ephrin (EPH) tyrosine kinase (such as Epha3,Ephb4), Ephrin B2 ligand, epidermal growth factor, epidermal growthfactor receptors (EGFR), epidermal growth factor receptor (EGFR) gene,Epigen, Epithelial cell adhesion molecule (EpCAM), Erb-b2 (v-erb-b2avian erythroblastic leukemia viral oncogene homolog 2) tyrosine kinasereceptor, Erb-b3 tyrosine kinase receptor, Erb-b4 tyrosine kinasereceptor, E-selectin, Estradiol 17 beta dehydrogenase, Estrogen receptor(such as alpha, beta), Estrogen related receptor, Eukaryotic translationinitiation factor 5A (EIF5A) gene, Exportin 1, Extracellular signalrelated kinase (such as 1, 2), Extracellular signal-regulated kinases(ERK), Factor (such as Xa, VIIa), farnesoid x receptor (FXR), Fasligand, Fatty acid synthase (FASN), Ferritin, FGF-2 ligand, FGF-5ligand, fibroblast growth factor (FGF, such as FGF1, FGF2, FGF4),Fibronectin, Fms-related tyrosine kinase 3 (Flt3), focal adhesion kinase(FAK, such as FAK2), folate hydrolase prostate-specific membrane antigen1 (FOLH1), Folate receptor (such as alpha), Folate, Folate transporter1, FYN tyrosine kinase, paired basic amino acid cleaving enzyme (FURIN),Beta-glucuronidase, Galactosyltransferase, Galectin-3, Ganglioside GD2,Glucocorticoid, glucocorticoid-induced TNFR-related protein GITRreceptor, Glutamate carboxypeptidase II, glutaminase, GlutathioneS-transferase P, glycogen synthase kinase (GSK, such as 3-beta),Glypican 3 (GPC3), gonadotropin-releaseing hormone (GNRH), Granulocytemacrophage colony stimulating factor (GM-CSF) receptor,Granulocyte-colony stimulating factor (GCSF) ligand, growth factorreceptor-bound protein 2 (GRB2), Grp78 (78 kDa glucose-regulatedprotein) calcium binding protein, molecular chaperone groEL2 gene, Heatshock protein (such as 27, 70, 90 alpha, beta), Heat shock protein gene,Heat stable enterotoxin receptor, Hedgehog protein, Heparanase,Hepatocyte growth factor, HERV-H LTR associating protein 2, Hexosekinase, Histamine H2 receptor, Histone methyltransferase (DOT1L),histone deacetylase (HDAC, such as 1, 2, 3, 6, 10, 11), Histone H1,Histone H3, HLA class I antigen (A-2 alpha), HLA class II antigen,Homeobox protein NANOG, HSPB1 gene, Human leukocyte antigen (HLA), Humanpapillomavirus (such as E6, E7) protein, Hyaluronic acid, Hyaluronidase,Hypoxia inducible factor-1 alpha (HIF1α), Imprinted Maternally ExpressedTranscript (H19) gene, mitogen-activated protein kinase kinase kinasekinase 1 (MAP4K1), tyrosine-protein kinase HCK, I-Kappa-B kinase (IKK,such as IKKbe), IL-1 alpha, IL-1 beta, IL-12, IL-12 gene, IL-15, IL-17,IL-2 gene, IL-2 receptor alpha subunit, IL-2, IL-3 receptor, IL-4, IL-6,IL-7, IL-8, immunoglobulin (such as G, G1, G2, K, M), Immunoglobulin Fcreceptor, Immunoglobulin gamma Fc receptor (such as I, III, IIIA),indoleamine 2,3-dioxygenase (IDO, such as IDO1), indoleamine pyrrole2,3-dioxygenase 1 inhibitor, insulin receptor, Insulin-like growthfactor (such as 1, 2), Integrin alpha-4/beta-1, integrin alpha-4/beta-7,Integrin alpha-5/beta-1, Integrin alpha-V/beta-3, Integrinalpha-V/beta-5, Integrin alpha-V/beta-6, Intercellular adhesion molecule1 (ICAM-1), interferon (such as alpha, alpha 2, beta, gamma), Interferoninducible protein absent in melanoma 2 (AIM2), interferon type Ireceptor, Interleukin 1 ligand, Interleukin 13 receptor alpha 2,interleukin 2 ligand, interleukin-1 receptor-associated kinase 4(IRAK4), Interleukin-2, Interleukin-29 ligand, isocitrate dehydrogenase(such as IDH1, IDH2), Janus kinase (JAK, such as JAK1, JAK2), Jun Nterminal kinase, kallikrein-related peptidase 3 (KLK3) gene, Killer cellIg like receptor, Kinase insert domain receptor (KDR), Kinesin-likeprotein KIF11, Kirsten rat sarcoma viral oncogene homolog (KRAS) gene,Kisspeptin (KiSS-1) receptor, KIT gene, v-kit Hardy-Zuckerman 4 felinesarcoma viral oncogene homolog (KIT) tyrosine kinase, lactoferrin,Lanosterol-14 demethylase, LDL receptor related protein-1, LeukotrieneA4 hydrolase, Listeriolysin, L-Selectin, Luteinizing hormone receptor,Lyase, lymphocyte activation gene 3 protein (LAG-3), Lymphocyte antigen75, Lymphocyte function antigen-3 receptor, lymphocyte-specific proteintyrosine kinase (LCK), Lymphotactin, Lyn (Lck/Yes novel) tyrosinekinase, lysine demethylases (such as KDM1, KDM2, KDM4, KDM5, KDM6,A/B/C/D), Lysophosphatidate-1 receptor, lysosomal-associated membraneprotein family (LAMP) gene, Lysyl oxidase homolog 2, lysyl oxidaseprotein (LOX), lysyl oxidase-like protein (LOXL, such as LOXL2),Hematopoietic Progenitor Kinase 1 (HPK1), Hepatocyte growth factorreceptor (MET) gene, macrophage colony-stimulating factor (MCSF) ligand,Macrophage migration inhibitory fact, MAGEC1 gene, MAGEC2 gene, Majorvault protein, MAPK-activated protein kinase (such as MK2), Mas-relatedG-protein coupled receptor, matrix metalloprotease (MMP, such as MMP2,MMP9), Mcl-1 differentiation protein, Mdm2 p53-binding protein, Mdm4protein, Melan-A (MART-1) melanoma antigen, Melanocyte protein Pmel 17,melanocyte stimulating hormone ligand, melanoma antigen family A3(MAGEA3) gene, Melanoma associated antigen (such as 1, 2, 3, 6),Membrane copper amine oxidase, Mesothelin, MET tyrosine kinase,Metabotropic glutamate receptor 1, Metalloreductase STEAP1 (sixtransmembrane epithelial antigen of the prostate 1), Metastin,methionine aminopeptidase-2, Methyltransferase, Mitochondrial 3 ketoacylCoA thiolase, mitogen-activate protein kinase (MAPK), mitogen-activatedprotein kinase (MEK, such as MEK1, MEK2), mTOR (mechanistic target ofrapamycin (serine/threonine kinase), mTOR complex (such as 1,2), mucin(such as 1, 5A, 16), mut T homolog (MTH, such as MTH1), Mycproto-oncogene protein, myeloid cell leukemia 1 (MCL1) gene,myristoylated alanine-rich protein kinase C substrate (MARCKS) protein,NAD ADP ribosyltransferase, natriuretic peptide receptor C, Neural celladhesion molecule 1, Neurokinin 1 (NK1) receptor, Neurokinin receptor,Neuropilin 2, NF kappa B activating protein, NIMA-related kinase 9(NEK9), Nitric oxide synthase, NK cell receptor, NK3 receptor, NKG2 A Bactivating NK receptor, Noradrenaline transporter, Notch (such asNotch-2 receptor, Notch-3 receptor, Notch-4 receptor), Nuclear erythroid2-related factor 2, Nuclear Factor (NF) kappa B, Nucleolin,Nucleophosmin, nucleophosmin-anaplastic lymphoma kinase (NPM-ALK), 2oxoglutarate dehydrogenase, 2,5-oligoadenylate synthetase,O-methylguanine DNA methyltransferase, Opioid receptor (such as delta),Ornithine decarboxylase, Orotate phosphoribosyltransferase, orphannuclear hormone receptor NR4A1, Osteocalcin, Osteoclast differentiationfactor, Osteopontin, OX-40 (tumor necrosis factor receptor superfamilymember 4 TNFRSF4, or CD134) receptor, P3 protein, p38 kinase, p38 MAPkinase, p53 tumor suppressor protein, Parathyroid hormone ligand,peroxisome proliferator-activated receptors (PPAR, such as alpha, delta,gamma), P-Glycoprotein (such as 1), phosphatase and tensin homolog(PTEN), phosphatidylinositol 3-kinase (PI3K), phosphoinositide-3 kinase(PI3K such as alpha, delta, gamma), phosphorylase kinase (PK), PKN3gene, placenta growth factor, platelet-derived growth factor (PDGF, suchas alpha, beta), Platelet-derived growth factor (PDGF, such as alpha,beta), Pleiotropic drug resistance transporter, Plexin B1, PLK1 gene,polo-like kinase (PLK), Polo-like kinase 1, Poly ADP ribose polymerase(PARP, such as PARP1, 2 and 3), Preferentially expressed antigen inmelanoma (PRAME) gene, Prenyl-binding protein (PrPB), Probabletranscription factor PML, Progesterone receptor, Programmed cell death 1(PD-1), Programmed cell death ligand 1 inhibitor (PD-L1), Prosaposin(PSAP) gene, Prostanoid receptor (EP4), prostate specific antigen,Prostatic acid phosphatase, proteasome, Protein E7, Proteinfarnesyltransferase, protein kinase (PK, such as A, B, C), proteintyrosine kinase, Protein tyrosine phosphatase beta, Proto-oncogeneserine/threonine-protein kinase (PIM, such as PIM-1, PIM-2, PIM-3),P-Selectin, Purine nucleoside phosphorylase, purinergic receptor P2Xligand gated ion channel 7 (P2X7), Pyruvate dehydrogenase (PDH),Pyruvate dehydrogenase kinase, Pyruvate kinase (PYK), 5-Alpha-reductase,Raf protein kinase (such as 1, B), RAF1 gene, Ras gene, Ras GTPase, RETgene, Ret tyrosine kinase receptor, retinoblastoma associated protein,retinoic acid receptor (such as gamma), Retinoid X receptor, Rheb (Rashomolog enriched in brain) GTPase, Rho (Ras homolog) associated proteinkinase 2, ribonuclease, Ribonucleotide reductase (such as M2 subunit),Ribosomal protein S6 kinase, RNA polymerase (such as I, II), Ron(Recepteur d'Origine Nantais) tyrosine kinase, ROS1 (ROS proto-oncogene1, receptor tyrosine kinase) gene, Rosi tyrosine kinase, Runt-relatedtranscription factor 3, Gamma-secretase, S100 calcium binding proteinA9, Sarco endoplasmic calcium ATPase, Second mitochondria-derivedactivator of caspases (SMAC) protein, Secreted frizzled relatedprotein-2, Semaphorin-4D, Serine protease, serine/threonine kinase(STK), serine/threonine-protein kinase (TBK, such as TBK1), signaltransduction and transcription (STAT, such as STAT-1, STAT-3, STAT-5),Signaling lymphocytic activation molecule (SLAM) family member 7,six-transmembrane epithelial antigen of the prostate (STEAP) gene, SLcytokine ligand, smoothened (SMO) receptor, Sodium iodide cotransporter,Sodium phosphate cotransporter 2B, Somatostatin receptor (such as 1, 2,3, 4, 5), Sonic hedgehog protein, Son of sevenless (SOS), Specificprotein 1 (Sp1) transcription factor, Sphingomyelin synthase,Sphingosine kinase (such as 1, 2), Sphingosine-1-phosphate receptor-1,spleen tyrosine kinase (SYK), SRC gene, Src tyrosine kinase, STAT3 gene,Steroid sulfatase, Stimulator of interferon genes (STING) receptor,stimulator of interferon genes protein, Stromal cell-derived factor 1ligand, SUMO (small ubiquitin-like modifier), Superoxide dismutase,Survivin protein, Synapsin 3, Syndecan-1, Synuclein alpha, T cellsurface glycoprotein CD28, tank-binding kinase (TBK), TATA box-bindingprotein-associated factor RNA polymerase I subunit B (TAF1B) gene,T-cell CD3 glycoprotein zeta chain, T-cell differentiation antigen CD6,T-cell immunoglobulin and mucin-domain containing-3 (TIM-3), T-cellsurface glycoprotein CD8, Tec protein tyrosine kinase, Tek tyrosinekinase receptor, telomerase, Telomerase reverse transcriptase (TERT)gene, Tenascin, TGF beta 2 ligand, Thrombopoietin receptor, Thymidinekinase, Thymidine phosphorylase, Thymidylate synthase, Thymosin (such asalpha 1), Thyroid hormone receptor, Thyroid stimulating hormonereceptor, Tissue factor, TNF related apoptosis inducing ligand, TNFR1associated death domain protein, TNF-related apoptosis-inducing ligand(TRAIL) receptor, TNFSF11 gene, TNFSF9 gene, Toll-like receptor (TLRsuch as 1-13), topoisomerase (such as I, II, III), Transcription factor,Transferase, Transferrin, Transforming growth factor (TGF, such as beta)kinase, Transforming growth factor TGF-β receptor kinase,Transglutaminase, Translocation associated protein, Transmembraneglycoprotein NMB, Trop-2 calcium signal transducer, trophoblastglycoprotein (TPBG) gene, Trophoblast glycoprotein, Tropomyosin receptorkinase (Trk) receptor (such as TrkA, TrkB, TrkC), Tryptophan5-hydroxylase, Tubulin, Tumor necrosis factor (TNF, such as alpha,beta), Tumor necrosis factor 13C receptor, tumor progression locus 2(TPL2), Tumor protein 53 (TP53) gene, Tumor suppressor candidate 2(TUSC2) gene, Tyrosinase, Tyrosine hydroxylase, tyrosine kinase (TK),Tyrosine kinase receptor, Tyrosine kinase with immunoglobulin-like andEGF-like domains (TIE) receptor, Tyrosine protein kinase ABL1 inhibitor,Ubiquitin, Ubiquitin carboxyl hydrolase isozyme L5, Ubiquitinthioesterase-14, Ubiquitin-conjugating enzyme E2I (UBE2I, UBC9), Urease,Urokinase plasminogen activator, Uteroglobin, Vanilloid VR1, Vascularcell adhesion protein 1, vascular endothelial growth factor receptor(VEGFR), V-domain Ig suppressor of T-cell activation (VISTA), VEGF-1receptor, VEGF-2 receptor, VEGF-3 receptor, VEGF-A, VEGF-B, Vimentin,Vitamin D3 receptor, Proto-oncogene tyrosine-protein kinase Yes, Wee-1protein kinase, Wilms' tumor antigen 1, Wilms' tumor protein, X-linkedinhibitor of apoptosis protein, Zinc finger protein transcription factoror any combination thereof.

Non-limiting examples of additional therapeutic agents may becategorized by their mechanism of action into, for example, thefollowing groups:

-   -   anti-metabolites/anti-cancer agents, such as pyrimidine analogs        floxuridine, capecitabine, cytarabine, CPX-351 (liposomal        cytarabine, daunorubicin), and TAS-118;    -   purine analogs, folate antagonists (such as pralatrexate), and        related inhibitors;    -   antiproliferative/antimitotic agents including natural products,        such as vinca alkaloids (vinblastine, vincristine) and        microtubule disruptors such as taxane (paclitaxel, docetaxel),        vinblastin, nocodazole, epothilones, vinorelbine (NAVELBINE®),        and epipodophyllotoxins (etoposide, teniposide);    -   DNA damaging agents, such as actinomycin, amsacrine, busulfan,        carboplatin, chlorambucil, cisplatin, cyclophosphamide        (CYTOXAN®), dactinomycin, daunorubicin, doxorubicin, epirubicin,        iphosphamide, melphalan, merchlorethamine, mitomycin C,        mitoxantrone, nitrosourea, procarbazine, taxol, Taxotere,        teniposide, etoposide, and triethylenethiophosphoramide;    -   DNA-hypomethylating agents, such as guadecitabine (SGI-110),        ASTX727;    -   antibiotics such as dactinomycin, daunorubicin, doxorubicin,        idarubicin, anthracyclines, mitoxantrone, bleomycins, plicamycin        (mithramycin);    -   enzymes such as L-asparaginase which systemically metabolizes        L-asparagine and deprives cells which do not have the capacity        to synthesize their own asparagine;    -   antiplatelet agents;    -   DNAi oligonucleotides targeting Bcl-2, such as PNT2258;    -   agents that activate or reactivate latent human immunodeficiency        virus (HIV), such as panobinostat and romidepsin;    -   asparaginase stimulators, such as crisantaspase (Erwinase®) and        GRASPA (ERY-001, ERY-ASP), calaspargase pegol;    -   pan-Trk, ROS1 and ALK inhibitors, such as entrectinib, TPX-0005;    -   anaplastic lymphoma kinase (ALK) inhibitors, such as alectinib,        ceritinib;    -   antiproliferative/antimitotic alkylating agents, such as        nitrogen mustard cyclophosphamide and analogs (melphalan,        chlorambucil, hexamethylmelamine, thiotepa), alkyl nitrosoureas        (carmustine) and analogs, streptozocin, and triazenes        (dacarbazine);    -   antiproliferative/antimitotic antimetabolites, such as folic        acid analogs (methotrexate);    -   platinum coordination complexes (cisplatin, oxiloplatinim, and        carboplatin), procarbazine, hydroxyurea, mitotane, and        aminoglutethimide;    -   hormones, hormone analogs (estrogen, tamoxifen, goserelin,        bicalutamide, and nilutamide), and aromatase inhibitors        (letrozole and anastrozole);    -   anticoagulants such as heparin, synthetic heparin salts, and        other inhibitors of thrombin;    -   fibrinolytic agents such as tissue plasminogen activator,        streptokinase, urokinase, aspirin, dipyridamole, ticlopidine,        and clopidogrel;    -   antimigratory agents;    -   antisecretory agents (breveldin);    -   immunosuppressives, such as tacrolimus, sirolimus, azathioprine,        and mycophenolate;    -   growth factor inhibitors, and vascular endothelial growth factor        inhibitors;    -   fibroblast growth factor inhibitors, such as FPA14;    -   anti-VEGFR antibodies, such as IMC-3C5, GNR-011, tanibirumab;    -   anti-VEGF/DDL4 antibodies, such as ABT-165;    -   anti-cadherins antibodies, such as HKT-288;    -   anti-CD70 antibodies, such as AMG-172; anti-leucine-rich repeat        containing 15 (LRRC15) antibodies, such as ABBV-085. ARGX-110;    -   angiotensin receptor blockers, nitric oxide donors;    -   antisense oligonucleotides, such as AEG35156, IONIS-KRAS-2.5Rx,        EZN-3042, RX-0201, IONIS-AR-2.5Rx, BP-100 (prexigebersen),        IONIS-STAT3-2.5Rx;    -   DNA interference oligonucleotides, such as PNT2258, AZD-9150;    -   anti-ANG-2 antibodies, such as MED13617, and LY3127804;    -   anti-ANG-1/ANG-2 antibodies, such as AMG-780;    -   anti-MET/EGFR antibodies, such as LY3164530;    -   anti-EGFR antibodies, such as ABT-414, AMG-595, necitumumab,        ABBV-221, depatuxizumab mafodotin (ABT-414), tomuzotuximab,        ABT-806, vectibix, modotuximab, RM-1929;    -   anti-CSF1R antibodies, such as emactuzumab, LY3022855, AMG-820,        FPA-008 (cabiralizumab);    -   anti-CD40 antibodies, such as RG7876, SEA-CD40, APX-005M,        ABBV-428;    -   anti-endoglin antibodies, such as TRC105 (carotuximab);    -   anti-CD45 antibodies, such as 131I-BC8 (lomab-B);    -   anti-HER3 antibodies, such as LJM716, GSK2849330;    -   anti-HER2 antibodies, such as margetuximab, MEDI4276, BAT-8001;    -   anti-HLA-DR antibodies, such as IMMU-114;    -   anti-IL-3 antibodies, such as JNJ-56022473;    -   anti-OX40 antibodies, such as MEDI6469, MEDI6383, MEDI0562        (tavolixizumab), MOXR0916, PF-04518600, RG-7888, GSK-3174998,        INCAGN1949, BMS-986178, GBR-8383, ABBV-368;    -   anti-EphA3 antibodies, such as KB-004;    -   anti-CD20 antibodies, such as obinutuzumab, IGN-002;    -   anti-CD20/CD3 antibodies, such as RG7828;    -   anti-CD37 antibodies, such as AGS67E, otlertuzumab (TRU-016);    -   anti-ENPP3 antibodies, such as AGS-16C3F;    -   anti-FGFR-3 antibodies, such as LY3076226, B-701;    -   anti-FGFR-2 antibodies, such as GAL-F2;    -   anti-C5 antibodies, such as ALXN-1210;    -   anti-CD27 antibodies, such as varlilumab (CDX-1127);    -   anti-TROP-2 antibodies, such as IMMU-132    -   anti-NKG2a antibodies, such as monalizumab;    -   anti-VISTA antibodies, such as HMBD-002;    -   anti-PVRIG antibodies, such as COM-701;    -   anti-EpCAM antibodies, such as VB4-845;    -   anti-BCMA antibodies, such as GSK-2857916    -   anti-CEA antibodies, such as RG-7813;    -   anti-cluster of differentiation 3 (CD3) antibodies, such as        MGD015;    -   anti-folate receptor alpha antibodies, such as IMGN853;    -   MCL-1 inhibitors, such as AMG-176, S-64315, and AZD-5991,        483-LM, A-1210477, UMI-77, JKY-5-037;    -   epha2 inhibitors, such as MM-310;    -   anti LAG-3 antibodies, such as relatlimab (ONO-4482), LAG-525,        MK-4280, REGN-3767;    -   raf kinase/VEGFR inhibitors, such as RAF-265;    -   polycomb protein (EED) inhibitors, such as MAK683;    -   anti-fibroblast activation protein (FAP)/IL-2R antibodies, such        as RG7461;    -   anti-fibroblast activation protein (FAP)/TRAIL-R2 antibodies,        such as RG7386;    -   anti-fucosyl-GM1 antibodies, such as BMS-986012;    -   p38 MAP kinase inhibitors, such as ralimetinib;    -   PRMT1 inhibitors, such as MS203;    -   Sphingosine kinase 2 (SK2) inhibitors, such as opaganib;    -   FLT3-ITD inhibitors, such as BCI-332;    -   Nuclear erythroid 2-related factor 2 stimulators, such as        omaveloxolone (RTA-408);    -   Tropomyosin receptor kinase (TRK) inhibitors, such as LOXO-195,        ONO-7579;    -   anti-ICOS antibodies, such as JTX-2011, GSK3359609;    -   anti-DR5 (TRAIL2) antibodies, such as DS-8273;    -   anti-GD2 antibodies, such as APN-301;    -   anti-interleukin-17 (IL-17) antibodies, such as CJM-112;    -   anti-carbonic anhydrase IX antibodies, such as TX-250;    -   anti-CD38-attenukine, such as TAK573;    -   anti-Mucin 1 antibodies, such as gatipotuzumab;    -   Mucin 1 inhibitors, such as GO-203-2C;    -   MARCKS protein inhibitors, such as BIO-11006;    -   Folate antagonists, such as arfolitixorin;    -   Galectin-3 inhibitors, such as GR-MD-02;    -   Phosphorylated P68 inhibitors, such as RX-5902;    -   CD95/TNF modulators, such as ofranergene obadenovec;    -   PI3K/Akt/mTOR inhibitors, such as ABTL-0812;    -   pan-PIM kinase inhibitors, such as INCB-053914;    -   IL-12 gene stimulators, such as EGEN-001, tavokinogene        telseplasmid;    -   Heat shock protein HSP90 inhibitors, such as TAS-116, PEN-866;    -   VEGF/HGF antagonists, such as MP-0250;    -   SYK tyrosine kinase/FLT3 tyrosine kinase inhibitors, such as        TAK-659;    -   SYK tyrosine kinase/JAK tyrosine kinase inhibitors, such as        ASN-002;    -   FLT3 tyrosine kinase inhibitor, such as FF-10101;    -   FLT3 tyrosine kinase agonist, such as CDX-301;    -   FLT3/MEK1 inhibitors, such as E-6201;    -   IL-24 antagonist, such as AD-IL24;    -   RIG-I agonists, such as RGT-100;    -   Aerolysin stimulators, such as topsalysin;    -   P-Glycoprotein 1 inhibitors, such as HM-30181A;    -   CSF-1 antagonists, such as ARRY-382, BLZ-945;    -   anti-Mesothelin antibodies, such as SEL-403;    -   Thymidine kinase stimulators, such as aglatimagene besadenovec;    -   Polo-like kinase 1 inhibitors, such as PCM-075;    -   TLR-7 agonists, such as TMX-101 (imiquimod);    -   NEDD8 inhibitors, such as pevonedistat (MLN-4924), TAS-4464;    -   Pleiotropic pathway modulators, such as avadomide (CC-122);    -   FoxM1 inhibitors, such as thiostrepton;    -   Anti-MUC1 antibodies, such as Mab-AR-20.5;    -   anti-CD38 antibodies, such as isatuximab, MOR-202;    -   UBA1 inhibitors, such as TAK-243;    -   Src tyrosine kinase inhibitors, such as VAL-201;    -   VDAC/HK inhibitors, such as VDA-1102;    -   BRAF/PI3K inhibitors, such as ASN-003;    -   Elf4a inhibitors, such as rohinitib, eFT226;    -   TP53 gene stimulators, such as ad-p53;    -   PD-L1/EGFR inhibitors, such as GNS-1480;    -   Retinoic acid receptor alpha (RARα) inhibitors, such as SY-1425;    -   SIRT3 inhibitors, such as YC8-02;    -   Stromal cell-derived factor 1 ligand inhibitors, such as        olaptesed pegol (NOX-A12);    -   IL-4 receptor modulators, such as MDNA-55;    -   Arginase-I stimulators, such as pegzilarginase;    -   Topoisomerase I inhibitor/hypoxia inducible factor-1 alpha        inhibitors, such as PEG-SN38 (firtecan pegol);    -   Hypoxia inducible factor-1 alpha inhibitors, such as PT-2977,        PT-2385;    -   CD122 agonists such as NKTR-214;    -   p53 tumor suppressor protein stimulators such as kevetrin;    -   Mdm4/Mdm2 p53-binding protein inhibitors, such as ALRN-6924;    -   kinesin spindle protein (KSP) inhibitors, such as filanesib        (ARRY-520);    -   CD80-fc fusion protein inhibitors, such as FPT-155;    -   Menin and mixed lineage leukemia (MLL) inhibitors such as        KO-539;    -   Liver x receptor agonists, such as RGX-104;    -   IL-10 agonists, such as AM-0010;    -   EGFR/ErbB-2 inhibitors, such as varlitinib;    -   VEGFR/PDGFR inhibitors, such as vorolanib;    -   IRAK4 inhibitors, such as CA-4948;    -   anti-TLR-2 antibodies, such as OPN-305;    -   Calmodulin modulators, such as CBP-501;    -   Glucocorticoid receptor antagonists, such as relacorilant        (CORT-125134);    -   Second mitochondria-derived activator of caspases (SMAC) protein        inhibitors, such as BI-891065;    -   Lactoferrin modulators, such as LTX-315;    -   Kit tyrosine kinase/PDGF receptor alpha antagonists such as        DCC-2618;    -   KIT inhibitors, such as PLX-9486;    -   Exportin 1 inhibitors, such as eltanexor;    -   EGFR/ErbB2/Ephb4 inhibitors, such as tesevatinib;    -   anti-CD33 antibodies, such as IMGN-779;    -   anti-KMA antibodies, such as MDX-1097;    -   anti-TIM-3 antibodies, such as TSR-022, LY-3321367, MBG-453;    -   anti-CD55 antibodies, such as PAT-SC1;    -   anti-PSMA antibodies, such as ATL-101;    -   anti-CD100 antibodies, such as VX-15;    -   anti-EPHA3 antibodies, such as fibatuzumab;    -   anti-Erbb antibodies, such as CDX-3379, HLX-02, seribantumab;    -   anti-APRIL antibodies, such as BION-1301;    -   Anti-Tigit antidbodies, such as BMS-986207, RG-6058;    -   CHST15 gene inhibitors, such as STNM-01;    -   RAS inhibitors, such as NEO-100;    -   Somatostatin receptor antagonist, such as OPS-201;    -   CEBPA gene stimulators, such as MTL-501;    -   DKK3 gene modulators, such as MTG-201;    -   p70s6k inhibitors, such as MSC2363318A;    -   methionine aminopeptidase 2 (MetAP2) inhibitors, such as M8891,        APL-1202;    -   arginine N-methyltransferase 5 inhibitors, such as GSK-3326595;    -   anti-programmed cell death protein 1 (anti-PD-1) antibodies,        such as nivolumab (OPDIVO®, BMS-936558, MDX-1106), pembrolizumab        (KEYTRUDA®, MK-3477, SCH-900475, lambrolizumab, CAS Reg. No.        1374853-91-4), pidilizumab, PF-06801591, BGB-A317, GLS-010        (WBP-3055), AK-103 (HX-008), MGA-012, BI-754091, REGN-2810        (cemiplimab), AGEN-2034, JS-001, JNJ-63723283, genolimzumab        (CBT-501), LZM-009, BCD-100, LY-3300054, SHR-1201, BAT-1306, and        anti-programmed death-ligand 1 (anti-PD-L1) antibodies such as        BMS-936559, atezolizumab (MPDL3280A), durvalumab (MED14736),        avelumab, CK-301, (MSB0010718C), MEDIO0680, CX-072, CBT-502,        PDR-001 (spartalizumab), TSR-042 (dostarlimab), JTX-4014,        BGB-A333, SHR-1316, CS-1001 (WBP-3155, KN-035, IBI-308, FAZ-053,        and MDX1105-01;    -   PD-L1/VISTA antagonists such as CA-170;    -   anti-PD-L1/TGFβ antibodies, such as M7824;    -   anti-transferrin antibodies, such as CX-2029;    -   anti-IL-8 (Interleukin-8) antibodies, such as HuMax-Inflam;    -   ATM (ataxia telangiectasia) inhibitors, such as AZD0156;    -   CHK1 inhibitors, such as GDC-0575, LY2606368 (prexasertib),        SRA737, RG7741 (CHK1/2);    -   CXCR4 antagonists, such as BL-8040, LY2510924, burixafor        (TG-0054), X4P-002, X4P-001-IO;    -   EXH2 inhibitors, such as GSK2816126;    -   HER2 inhibitors, such as neratinib, tucatinib (ONT-380);    -   KDM1 inhibitors, such as ORY-1001, IMG-7289, INCB-59872,        GSK-2879552;    -   CXCR2 antagonists, such as AZD-5069;    -   GM-CSF antibodies, such as lenzilumab;    -   DNA dependent protein kinase inhibitors, such as MSC2490484A        (nedisertib), VX-984, AsiDNA (DT-01);    -   protein kinase C (PKC) inhibitors, such as LXS-196,        sotrastaurin;    -   Selective estrogen receptor downregulators (SERD), such as        fulvestrant (Faslodex®), RG6046, RG6047, elacestrant (RAD-1901)        and AZD9496;    -   Selective estrogen receptor covalent antagonists (SERCAs), such        as H3B-6545;    -   selective androgen receptor modulator (SARM), such as GTX-024,        darolutamide;    -   transforming growth factor-beta (TGF-beta) kinase antagonists,        such as galunisertib;    -   anti-transforming growth factor-beta (TGF-beta) antibodies, such        as LY3022859, NIS793, XOMA 089;    -   bispecific antibodies, such as MM-141 (IGF-1/ErbB3), MM-111        (Erb2/Erb3), JNJ-64052781 (CD19/CD3), PRS-343 (CD-137/HER2),        AFM26 (BCMA/CD16A), JNJ-61186372 (EGFR/cMET), AMG-211 (CEA/CD3),        RG7802 (CEA/CD3), ERY-974 (CD3/GPC3) vancizumab        (angiopoietins/VEGF), PF-06671008 (Cadherins/CD3), AFM-13        (CD16/CD30), APVO436 (CD123/CD3), flotetuzumab (CD123/CD3),        REGN-1979 (CD20/CD3), MCLA-117 (CD3/CLEC12A), MCLA-128        (HER2/HER3), JNJ-0819, JNJ-7564 (CD3/heme), AMG-757 (DLL3-CD3),        MGD-013 (PD-1/LAG-3), AK-104 (CTLA-4/PD-1), AMG-330 (CD33/CD3),        AMG-420 (BCMA/CD3), BI-836880 (VEFG/ANG2), JNJ-63709178        (CD123/CD3), MGD-007 (CD3/gpA33), MGD-009 (CD3/B7H3);    -   Mutant selective EGFR inhibitors, such as PF-06747775, EGF816        (nazartinib), ASP8273, ACEA-0010, BI-1482694;    -   Anti-GITR (glucocorticoid-induced tumor necrosis factor        receptor-related protein) antibodies, such as MEDI1873, FPA-154,        INCAGN-1876, TRX-518, BMS-986156, MK-1248, GWN-323;    -   anti-delta-like protein ligand 3 (DDL3) antibodies, such as        rovalpituzumab tesirine;    -   anti-clusterin antibodies, such as AB-16B5;    -   anti-Ephrin-A4 (EFNA4) antibodies, such as PF-06647263;    -   anti-RANKL antibodies, such as denosumab;    -   anti-mesothelin antibodies, such as BMS-986148, Anti-MSLN-MMAE;    -   anti-sodium phosphate cotransporter 2B (NaP2B) antibodies, such        as lifastuzumab    -   anti-c-Met antibodies, such as ABBV-399;    -   Adenosine A2A receptor antagonists, such as CPI-444, AZD-4635,        preladenant, PBF-509;    -   Alpha-ketoglutarate dehydrogenase (KGDH) inhibitors, such as        CPI-613;    -   XPO1 inhibitors, such as selinexor (KPT-330);    -   Isocitrate dehydrogenase 2 (IDH2) inhibitors, such as enasidenib        (AG-221);    -   IDH1 inhibitors such as AG-120, and AG-881 (IDH1 and IDH2),        IDH-305, BAY-1436032;    -   interleukin-3 receptor (IL-3R) modulators, such as SL-401;    -   Arginine deiminase stimulators, such as pegargiminase        (ADI-PEG-20);    -   antibody-drug conjugates, such as MLN0264 (anti-GCC, guanylyl        cyclase C), T-DM1 (trastuzumab emtansine, Kadcycla),        milatuzumab-doxorubicin (hCD74-DOX), brentuximab vedotin,        DCDT2980S, polatuzumab vedotin, SGN-CD70A, SGN-CD19A, inotuzumab        ozogamicin, lorvotuzumab mertansine, SAR3419, isactuzumab        govitecan, enfortumab vedotin (ASG-22ME), ASG-15ME, DS-8201        ((trastuzumab deruxtecan), 225Ac-lintuzumab, U3-1402,        177Lu-tetraxetan-tetuloma, tisotumab vedotin, anetumab        ravtansine, CX-2009, SAR-566658, W-0101, polatuzumab vedotin,        ABBV-085;    -   claudin-18 inhibitors, such as claudiximab;    -   β-catenin inhibitors, such as CWP-291;    -   anti-CD73 antibodies, such as MEDI-9447 (oleclumab), CPX-006,        IPH-53, BMS-986179;    -   CD73 antagonists, such as AB-680, PSB-12379, PSB-12441,        PSB-12425;    -   CD39/CD73 antagonists, such as PBF-1662;    -   chemokine receptor 2 (CCR) inhibitors, such as PF-04136309,        CCX-872, BMS-813160 (CCR2/CCR5)    -   thymidylate synthase inhibitors, such as ONX-0801;    -   ALK/ROS1 inhibtors, such as lorlatinib;    -   tankyrase inhibitors, such as G007-LK;    -   Mdm2 p53-binding protein inhibitors, such as CMG-097, HDM-201;    -   c-PIM inhibitors, such as PIM447;    -   BRAF inhibitors, such as dabrafenib, vemurafenib, encorafenib        (LGX818), PLX8394;    -   sphingosine kinase-2 (SK2) inhibitors, such as Yeliva®        (ABC294640);    -   cell cycle inhibitors, such as selumetinib (MEK1/2), and        sapacitabine;    -   AKT inhibitors such as MK-2206, ipatasertib, afuresertib,        AZD5363, and ARQ-092, capivasertib, triciribine;    -   anti-CTLA-4 (cytotoxic T-lymphocyte protein-4) inhibitors, such        as tremelimumab, AGEN-1884, BMS-986218;    -   c-MET inhibitors, such as AMG-337, savolitinib, tivantinib        (ARQ-197), capmatinib, and tepotinib, ABT-700, AG213, AMG-208,        JNJ-38877618 (OMO-1), merestinib, HQP-8361;    -   c-Met/VEGFR inhibitors, such as BMS-817378, TAS-115;    -   c-Met/RON inhibitors, such as BMS-777607;    -   BRAF/EGFR inhibitors, such as BGB-283;    -   bcr/abl inhibitors, such as rebastinib, asciminib;    -   MNK1/MNK2 inhibitors, such as eFT-508;    -   mTOR inhibitor/cytochrome P450 3A4 stimulators, such as TYME-88    -   lysine-specific demethylase-1 (LSD1) inhibitors, such as        CC-90011;    -   Pan-RAF inhibitors, such as LY3009120, LXH254, TAK-580;    -   Raf/MEK inhibitors, such as RG7304;    -   CSF1R/KIT and FLT3 inhibitors, such as pexidartinib (PLX3397);    -   kinase inhibitors, such as vandetanib;    -   E selectin antagonists, such as GMI-1271;    -   differentiation inducers, such as tretinoin;    -   epidermal growth factor receptor (EGFR) inhibitors, such as        osimertinib (AZD-9291);    -   topoisomerase inhibitors, such as doxorubicin, daunorubicin,        dactinomycin, eniposide, epirubicin, etoposide, idarubicin,        irinotecan, mitoxantrone, pixantrone, sobuzoxane, topotecan,        irinotecan, MM-398 (liposomal irinotecan), vosaroxin and        GPX-150, aldoxorubicin, AR-67, mavelertinib, AST-2818, avitinib        (ACEA-0010), irofulven (MGI-114);    -   corticosteroids, such as cortisone, dexamethasone,        hydrocortisone, methylprednisolone, prednisone, prednisolone;    -   growth factor signal transduction kinase inhibitors;    -   nucleoside analogs, such as DFP-10917;    -   Axl inhibitors, such as BGB-324 (bemcentinib), SLC-0211;    -   BET inhibitors, such as INCB-054329, INCB057643, TEN-010,        AZD-5153, ABT-767, BMS-986158, CC-90010, GSK525762 (molibresib),        NHWD-870, ODM-207, GSK-2820151, GSK-1210151A, ZBC246, ZBC260,        ZEN3694, FT-1101, RG-6146, CC-90010, mivebresib, BI-894999,        PLX-2853, PLX-51107, CPI-0610, GS-5829;    -   PARP inhibitors, such as olaparib, rucaparib, veliparib,        talazoparib, ABT-767, BGB-290;    -   Proteasome inhibitors, such as ixazomib, carfilzomib        (Kyprolis®), marizomib;    -   Glutaminase inhibitors, such as CB-839;    -   Vaccines, such as peptide vaccine TG-01 (RAS), GALE-301,        GALE-302, nelipepimut-s, SurVaxM, DSP-7888, TPIV-200, PVX-410,        VXL-100, DPX-E7, ISA-101, 6MHP, OSE-2101, galinpepimut-S,        SVN53-67/M57-KLH, IMU-131; bacterial vector vaccines such as        CRS-207/GVAX, axalimogene filolisbac (ADXS11-001); adenovirus        vector vaccines such as nadofaragene firadenovec; autologous        Gp96 vaccine; dendritic cells vaccines, such as CVactm,        stapuldencel-T, eltrapuldencel-T, SL-701, BSK01™,        rocapuldencel-T (AGS-003), DCVAC, CVac™, stapuldencel-T,        eltrapuldencel-T, SL-701, BSK01™, ADXS31-142; oncolytic vaccines        such as, talimogene laherparepvec, pexastimogene devacirepvec,        GL-ONC1, MG1-MA3, parvovirus H-1, ProstAtak, enadenotucirev,        MG1MA3, ASN-002 (TG-1042); therapeutic vaccines, such as        CVAC-301, CMP-001, PF-06753512, VBI-1901, TG-4010, ProscaVax™;        tumor cell vaccines, such as Vigil® (IND-14205), Oncoquest-L        vaccine; live attenuated, recombinant, serotype 1 poliovirus        vaccine, such as PVS-RIPO; Adagloxad simolenin; MEDI-0457;        DPV-001 a tumor-derived, autophagosome enriched cancer vaccine;        RNA vaccines such as CV-9209, LV-305; DNA vaccines, such as        MEDI-0457, MVI-816, INO-5401; modified vaccinia virus Ankara        vaccine expressing p53, such as MVA-p53; DPX-Survivac; BriaVax™;        GI-6301; GI-6207; GI-4000;    -   anti-DLL4 (delta like ligand 4) antibodies, such as demcizumab;    -   STAT-3 inhibitors, such as napabucasin (BBI-608);    -   ATPase p97 inhibitors, such as CB-5083;    -   smoothened (SMO) receptor inhibitors, such as Odomzo®        (sonidegib, formerly LDE-225), LEQ506, vismodegib (GDC-0449),        BMS-833923, glasdegib (PF-04449913), LY2940680, and        itraconazole;    -   interferon alpha ligand modulators, such as interferon alpha-2b,        interferon alpha-2a biosimilar (Biogenomics), ropeginterferon        alfa-2b (AOP-2014, P-1101, PEG IFN alpha-2b), Multiferon        (Alfanative, Viragen), interferon alpha 1b, Roferon-A (Canferon,        Ro-25-3036), interferon alfa-2a follow-on biologic        (Biosidus)(Inmutag, Inter 2A), interferon alfa-2b follow-on        biologic (Biosidus-Bioferon, Citopheron, Ganapar, Beijing Kawin        Technology—Kaferon), Alfaferone, pegylated interferon alpha-1b,        peginterferon alfa-2b follow-on biologic (Amega), recombinant        human interferon alpha-1b, recombinant human interferon        alpha-2a, recombinant human interferon alpha-2b, veltuzumab-IFN        alpha 2b conjugate, Dynavax (SD-101), and interferon alfa-nl        (Humoferon, SM-10500, Sumiferon);    -   interferon gamma ligand modulators, such as interferon gamma        (OH-6000, Ogamma 100);    -   IL-6 receptor modulators, such as tocilizumab, siltuximab,        AS-101 (CB-06-02, IVX-Q-101);    -   Telomerase modulators, such as, tertomotide (GV-1001, HR-2802,        Riavax) and imetelstat (GRN-163, JNJ-63935937);    -   DNA methyltransferases inhibitors, such as temozolomide        (CCRG-81045), decitabine, guadecitabine (S-110, SGI-110),        KRX-0402, RX-3117, RRx-001, and azacitidine;    -   DNA gyrase inhibitors, such as pixantrone and sobuzoxane;    -   Bcl-2 family protein inhibitors, such as ABT-263, venetoclax        (ABT-199), ABT-737, and AT-101;    -   Notch inhibitors, such as LY3039478 (crenigacestat), tarextumab        (anti-Notch2/3), BMS-906024;    -   anti-myostatin inhibitors, such as landogrozumab;    -   hyaluronidase stimulators, such as PEGPH-20;    -   Wnt pathway inhibitors, such as SM-04755, PRI-724, WNT-974;    -   gamma-secretase inhibitors, such as PF-03084014, MK-0752,        RO-4929097;    -   Grb-2 (growth factor receptor bound protein-2) inhibitors, such        as BP1001;    -   TRAIL pathway-inducing compounds, such as ONC201, ABBV-621;    -   Focal adhesion kinase inhibitors, such as VS-4718, defactinib,        GSK2256098;    -   hedgehog inhibitors, such as saridegib, sonidegib (LDE225),        glasdegib and vismodegib;    -   Aurora kinase inhibitors, such as alisertib (MLN-8237), and        AZD-2811, AMG-900, barasertib, ENMD-2076;    -   HSPB1 modulators (heat shock protein 27, HSP27), such as        brivudine, apatorsen;    -   ATR inhibitors, such as BAY-937, AZD6738, AZD6783, VX-803,        VX-970 (berzosertib) and VX-970;    -   mTOR inhibitors, such as sapanisertib and vistusertib (AZD2014),        ME-344;    -   mTOR/PI3K inhibitors, such as gedatolisib, GSK2141795,        omipalisib, RG6114;    -   Hsp90 inhibitors, such as AUY922, onalespib (AT13387), SNX-2112,        SNX5422;    -   Murine double minute (mdm2) oncogene inhibitors, such as        DS-3032b, RG7775, AMG-232, HDM201, and idasanutlin (RG7388);    -   CD137 agonists, such as urelumab, utomilumab (PF-05082566);    -   STING agonists, such as ADU-S100 (MIW-815), SB-11285, MK-1454,        SR-8291, AdVCA0848, GSK-532, SYN-STING, MSA-1, SR-8291;    -   FGFR inhibitors, such as FGF-401, INCB-054828, BAY-1163877,        AZD4547, JNJ-42756493, LY2874455, Debio-1347;    -   fatty acid synthase (FASN) inhibitors, such as TVB-2640;    -   Anti-KIR monoclonal antibodies, such as lirilumab (IPH-2102),        IPH-4102;    -   Antigen CD19 inhibitors, such as MOR208, MEDI-551, AFM-11,        inebilizumab;    -   CD44 binders, such as A6;    -   protein phosphatease 2A (PP2A) inhibitors, such as LB-100;    -   CYP17 inhibitors, such as seviteronel (VT-464), ASN-001,        ODM-204, CFG920, abiraterone acetate;    -   RXR agonists, such as IRX4204;    -   hedgehog/smoothened (hh/Smo) antagonists, such as taladegib,        patidegib;    -   complement C3 modulators, such as Imprime PGG;    -   IL-15 agonists, such as ALT-803, NKTR-255, and hetIL-15;    -   EZH2 (enhancer of zeste homolog 2) inhibitors, such as        tazemetostat, CPI-1205, GSK-2816126;    -   Oncolytic viruses, such as pelareorep, CG-0070, MV-NIS therapy,        HSV-1716, DS-1647, VCN-01, ONCOS-102, TBI-1401, tasadenoturev        (DNX-2401), vocimagene amiretrorepvec, RP-1, CVA21, Celyvir,        LOAd-703, OBP-301;    -   DOT1L (histone methyltransferase) inhibitors, such as        pinometostat (EPZ-5676);    -   toxins such as Cholera toxin, ricin, Pseudomonas exotoxin,        Bordetella pertussis adenylate cyclase toxin, diphtheria toxin,        and caspase activators;    -   DNA plasmids, such as BC-819    -   PLK inhibitors of PLK 1, 2, and 3, such as volasertib (PLK1);    -   WEE1 inhibitors, such as AZD1775 (adavosertib);    -   Rho kinase (ROCK) inhibitors, such as AT13148, KD025;    -   ERK inhibitors, such as GDC-0994, LY3214996, MK-8353;    -   IAP inhibitors, such as ASTX660, debio-1143, birinapant,        APG-1387, LCL-161;    -   RNA polymerase inhibitors, such has lurbinectedin (PM-1183),        CX-5461;    -   Tubulin inhibitors, such as PM-184, BAL-101553 (lisavanbulin),        and OXI-4503, fluorapacin (AC-0001), plinabulin;    -   Toll-like receptor 4 (TL4) agonists, such as G100, GSK1795091,        and PEPA-10;    -   Elongation factor 1 alpha 2 inhibitors, such as plitidepsin;    -   CD95 inhibitors, such as APG-101, APO-010, asunercept;    -   WT1 inhibitors, such as DSP-7888;    -   splicing factor 3B subunit1 (SF3B1) inhibitors, such as H3B-8800    -   PDGFR alpha/KIT mutant-specific inhibitors such as BLU-285;    -   SHP-2 inhibitors, such as TNO155 (SHP-099), RMC-4550; and    -   retinoid Z receptor gamma (RORγ) agonists, such as LYC-55716.

In some embodiments, provided herein are methods of treating orpreventing a hyperproliferative disorder or cancer in a human or animalhaving or at risk of having the hyperproliferative disorder or cancer isprovided, comprising administering to the human or animal atherapeutically effective amount of a compound of the presentdisclosure, or a pharmaceutically acceptable salt thereof, incombination with a therapeutically effective amount of one or more(e.g., one, two, three, one or two, or one to three) additionaltherapeutic agents selected from the group consisting of apoptosissignal-regulating kinase (ASK) inhibitors; Bruton's tyrosine kinase(BTK) inhibitors; cluster of differentiation 47 (CD47) inhibitors;cyclin-dependent kinase (CDK) inhibitors; discoidin domain receptor(DDR) inhibitors; histone deacetylase (HDAC) inhibitors;indoleamine-pyrrole-2,3-dioxygenase (IDO1) inhibitors; Janus kinase(JAK) inhibitors; lysyl oxidase-like protein (LOXL) inhibitors; matrixmetalloprotease (MMP) inhibitors; mitogen-activated protein kinase (MEK)inhibitors; phosphatidylinositol 3-kinase (PI3K) inhibitors; spleentyrosine kinase (SYK) inhibitors; toll-like receptor 8 (TLR8)inhibitors; toll-like receptor 9 (TLR9) inhibitors; tyrosine-kinaseinhibitors (TKIs), and any combination thereof, or a pharmaceuticallyacceptable salt thereof. Non-limiting examples include:

-   -   Apoptosis Signal-Regulating Kinase (ASK) Inhibitors: ASK        inhibitors include ASK1 inhibitors. Examples of ASK1 inhibitors        include, but are not limited to, those described in WO        2011/008709 (Gilead Sciences) and WO 2013/112741 (Gilead        Sciences);    -   Bruton's Tyrosine Kinase (BTK) Inhibitors: Examples of BTK        inhibitors include, but are not limited to,        (S)-6-amino-9-(1-(but-2-ynoyl)pyrrolidin-3-yl)-7-(4-phenoxyphenyl)-7H-purin-8(9H)-one,        acalabrutinib (ACP-196), BGB-3111, CB988, HM71224, ibrutinib,        M-2951 (evobrutinib), M7583, tirabrutinib (ONO-4059), PRN-1008,        spebrutinib (CC-292), TAK-020, vecabrutinib, ARQ-531, SHR-1459,        DTRMWXHS-12, TAS-5315;    -   Cluster of Differentiation 47 (CD47) inhibitors: Examples of        CD47 inhibitors include, but are not limited to anti-CD47 mAbs        (Vx-1004), anti-human CD47 mAbs (CNTO-7108), CC-90002,        CC-90002-ST-001, humanized anti-CD47 antibody (Hu5F9-G4),        NI-1701, NI-1801, RCT-1938, and TTI-621;    -   Cyclin-dependent Kinase (CDK) Inhibitors: CDK inhibitors include        inhibitors of CDK 1, 2, 3, 4, 6, 7 and 9, such as abemaciclib,        alvocidib (HMR-1275, flavopiridol), AT-7519, dinaciclib,        ibrance, FLX-925, LEE001, palbociclib, ribociclib, rigosertib,        selinexor, UCN-01, SY1365, CT-7001, SY-1365, G1T38, milciclib,        trilaciclib, and TG-02;    -   Discoidin Domain Receptor (DDR) Inhibitors: DDR inhibitors        include inhibitors of DDR1 and/or DDR2. Examples of DDR        inhibitors include, but are not limited to, those disclosed in        WO 2014/047624 (Gilead Sciences), US 2009-0142345 (Takeda        Pharmaceutical), US 2011-0287011 (Oncomed Pharmaceuticals), WO        2013/027802 (Chugai Pharmaceutical), and WO 2013/034933        (Imperial Innovations);    -   Histone Deacetylase (HDAC) Inhibitors: Examples of HDAC        inhibitors include, but are not limited to, abexinostat,        ACY-241, AR-42, BEBT-908, belinostat, CKD-581, CS-055        (HBI-8000), CUDC-907 (fimepinostat), entinostat, givinostat,        mocetinostat, panobinostat, pracinostat, quisinostat        (JNJ-26481585), resminostat, ricolinostat, SHP-141, valproic        acid (VAL-001), vorinostat, tinostamustine, remetinostat,        entinostat;    -   Indoleamine-pyrrole-2,3-dioxygenase (IDO1) inhibitors: Examples        of IDO1 inhibitors include, but are not limited to, BLV-0801,        epacadostat, F-001287, GBV-1012, GBV-1028, GDC-0919, indoximod,        NKTR-218, NLG-919-based vaccine, PF-06840003,        pyranonaphthoquinone derivatives (SN-35837), resminostat,        SBLK-200802, BMS-986205, and shIDO-ST, EOS-200271, KHK-2455,        LY-3381916;    -   Janus Kinase (JAK) Inhibitors: JAK inhibitors inhibit JAK1,        JAK2, and/or JAK3. Examples of JAK inhibitors include, but are        not limited to, AT9283, AZD1480, baricitinib, BMS-911543,        fedratinib, filgotinib (GLPG0634), gandotinib (LY2784544),        INCB039110 (itacitinib), lestaurtinib, momelotinib (CYT0387),        NS-018, pacritinib (SB1518), peficitinib (ASP015K), ruxolitinib,        tofacitinib (formerly tasocitinib), INCB052793, and XL019;    -   Lysyl Oxidase-Like Protein (LOXL) Inhibitors: LOXL inhibitors        include inhibitors of LOXL1, LOXL2, LOXL3, LOXL4, and/or LOXL5.        Examples of LOXL inhibitors include, but are not limited to, the        antibodies described in WO 2009/017833 (Arresto Biosciences).        Examples of LOXL2 inhibitors include, but are not limited to,        the antibodies described in WO 2009/017833 (Arresto        Biosciences), WO 2009/035791 (Arresto Biosciences), and WO        2011/097513 (Gilead Biologics);    -   Matrix Metalloprotease (MVP) Inhibitors: MMP inhibitors include        inhibitors of MMP1 through 10. Examples of MMP9 inhibitors        include, but are not limited to, marimastat (BB-2516),        cipemastat (Ro 32-3555), GS-5745 (andecaliximab) and those        described in WO 2012/027721 (Gilead Biologics);    -   Mitogen-activated Protein Kinase (MEK) Inhibitors: MEK        inhibitors include antroquinonol, binimetinib, cobimetinib        (GDC-0973, XL-518), MT-144, selumetinib (AZD6244), sorafenib,        trametinib (GSK1120212), uprosertib+trametinib, PD-0325901,        pimasertib, LTT462, AS703988, CC-90003, refametinib;    -   Phosphatidylinositol 3-kinase (PI3K) Inhibitors: PI3K inhibitors        include inhibitors of PI3Kγ, PI3Kδ, PI3Kβ, PI3Kα, and/or        pan-PI3K. Examples of PI3K inhibitors include, but are not        limited to, ACP-319, AEZA-129, AMG-319, AS252424, AZD8186, BAY        10824391, BEZ235, buparlisib (BKM120), BYL719 (alpelisib),        CH5132799, copanlisib (BAY 80-6946), duvelisib, GDC-0032,        GDC-0077, GDC-0941, GDC-0980, GSK2636771, GSK2269557, idelalisib        (Zydelig®), INCB50465, IPI-145, IPI-443, IPI-549, KAR4141,        LY294002, LY3023414, MLN1117, OXY111A, PA799, PX-866, RG7604,        rigosertib, RP5090, RP6530, SRX3177, taselisib, TG100115,        TGR-1202 (umbralisib), TGX221, WX-037, X-339, X-414, XL147        (SAR245408), XL499, XL756, wortmannin, ZSTK474, and the        compounds described in WO 2005/113556 (ICOS), WO 2013/052699        (Gilead Calistoga), WO 2013/116562 (Gilead Calistoga), WO        2014/100765 (Gilead Calistoga), WO 2014/100767 (Gilead        Calistoga), and WO 2014/201409 (Gilead Sciences);    -   Spleen Tyrosine Kinase (SYK) Inhibitors: Examples of SYK        inhibitors include, but are not limited to,        6-(1H-indazol-6-yl)-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine,        BAY-61-3606, cerdulatinib (PRT-062607), entospletinib,        fostamatinib (R788), HMPL-523, NVP-QAB 205 AA, R112, R343,        tamatinib (R406), and those described in U.S. Pat. No. 8,450,321        (Gilead Connecticut) and those described in U.S. 2015/0175616;    -   Toll-like receptor 8 (TLR8) inhibitors: Examples of TLR8        inhibitors include, but are not limited to, E-6887, IMO-4200,        IMO-8400, IMO-9200, MCT-465, MEDI-9197, motolimod, resiquimod,        VTX-1463, and VTX-763;    -   Toll-like receptor 9 (TLR9) inhibitors: Examples of TLR9        inhibitors include, but are not limited to, AST-008, IMO-2055,        IMO-2125, lefitolimod, litenimod, MGN-1601, and PUL-042; and    -   Tyrosine-kinase Inhibitors (TKIs): TKIs may target epidermal        growth factor receptors (EGFRs) and receptors for fibroblast        growth factor (FGF), platelet-derived growth factor (PDGF), and        vascular endothelial growth factor (VEGF). Examples of TKIs        include, but are not limited to, afatinib, ARQ-087        (derazantinib), asp5878, AZD3759, AZD4547, bosutinib,        brigatinib, cabozantinib, cediranib, crenolanib, dacomitinib,        dasatinib, dovitinib, E-6201, erdafitinib, erlotinib, gefitinib,        gilteritinib (ASP-2215), FP-1039, HM61713, icotinib, imatinib,        KX2-391 (Src), lapatinib, lestaurtinib, lenvatinib, midostaurin,        nintedanib, ODM-203, osimertinib (AZD-9291), ponatinib,        poziotinib, quizartinib, radotinib, rociletinib, sulfatinib        (HMPL-012), sunitinib, tivoanib, and TH-4000, MEDI-575        (anti-PDGFR antibody).

As used herein, the term “chemotherapeutic agent” or “chemotherapeutic”(or “chemotherapy” in the case of treatment with a chemotherapeuticagent) is meant to encompass any non-proteinaceous (i.e., non-peptidic)chemical compound useful in the treatment of cancer. Examples ofchemotherapeutic agents include but are not limited to: alkylatingagents such as thiotepa and cyclophosphamide (CYTOXAN®); alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridinessuch as benzodepa, carboquone, meturedepa, and uredepa; ethyleniminesand methylamelamines including altretamine, triethylenemelamine,triethylenephosphoramide, triethylenethiophosphoramide, andtrimemylolomelamine; acetogenins, especially bullatacin andbullatacinone; a camptothecin, including synthetic analog topotecan;bryostatin, callystatin; CC-1065, including its adozelesin, carzelesin,and bizelesin synthetic analogs; cryptophycins, particularlycryptophycin 1 and cryptophycin 8; dolastatin; duocarmycin, includingthe synthetic analogs KW-2189 and CBI-TMI; eleutherobin; 5-azacytidine;pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such aschlorambucil, chlomaphazine, cyclophosphamide, glufosfamide,evofosfamide, bendamustine, estramustine, ifosfamide, mechlorethamine,mechlorethamine oxide hydrochloride, melphalan, novembichin,phenesterine, prednimustine, trofosfamide, and uracil mustard;nitrosoureas such as carmustine, chlorozotocin, foremustine, lomustine,nimustine, and ranimustine; antibiotics such as the enediyne antibiotics(e.g., calicheamicin, especially calicheamicin gammall and calicheamicinphiI1), dynemicin including dynemicin A, bisphosphonates such asclodronate, an esperamicin, neocarzinostatin chromophore and relatedchromoprotein enediyne antibiotic chromomophores, aclacinomycins,actinomycin, authramycin, azaserine, bleomycins, cactinomycin,carabicin, carrninomycin, carzinophilin, chromomycins, dactinomycin,daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin(including morpholino-doxorubicin, cyanomorpholino-doxorubicin,2-pyrrolino-doxorubicin, and deoxydoxorubicin), epirubicin, esorubicin,idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolicacid, nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin,quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,ubenimex, zinostatin, and zorubicin; anti-metabolites such asmethotrexate and 5-fluorouracil (5-FU); folic acid analogs such asdemopterin, methotrexate, pteropterin, and trimetrexate; purine analogssuch as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine;pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine,carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, andfloxuridine; androgens such as calusterone, dromostanolone propionate,epitiostanol, mepitiostane, and testolactone; anti-adrenals such asaminoglutethimide, mitotane, and trilostane; folic acid replinisherssuch as frolinic acid; radiotherapeutic agents such as Radium-223;trichothecenes, especially T-2 toxin, verracurin A, roridin A, andanguidine; taxoids such as paclitaxel (TAXOL®), abraxane, docetaxel(TAXOTERE®), cabazitaxel, BIND-014, tesetaxel; platinum analogs such ascisplatin and carboplatin, NC-6004 nanoplatin; aceglatone;aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine;hestrabucil; bisantrene; edatraxate; defofamine; demecolcine;diaziquone; elformthine; elliptinium acetate; an epothilone; etoglucid;gallium nitrate; hydroxyurea; lentinan; leucovorin; lonidamine;maytansinoids such as maytansine and ansamitocins; mitoguazone;mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin;losoxantrone; fluoropyrimidine; folinic acid; podophyllinic acid;2-ethylhydrazide; procarbazine; polysaccharide-K (PSK); razoxane;rhizoxin; sizofiran; spirogermanium; tenuazonic acid; trabectedin,triaziquone; 2,2′,2″-tricUorotriemylamine; urethane; vindesine;dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman;gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiopeta;chlorambucil; gemcitabine (GEMZAR®); 6-thioguanine; mercaptopurine;methotrexate; vinblastine; platinum; etoposide (VP-16); ifosfamide;mitroxantrone; vancristine; vinorelbine (NAVELBINE®); novantrone;teniposide; edatrexate; daunomycin; aminopterin; xeoloda; ibandronate;CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylomithine (DFMO);retinoids such as retinoic acid; capecitabine; NUC-1031; FOLFIRI(fluorouracil, leucovorin, and irinotecan); and pharmaceuticallyacceptable salts, acids, or derivatives of any of the above.

Also included in the definition of “chemotherapeutic agent” areanti-hormonal agents such as anti-estrogens and selective estrogenreceptor modulators (SERMs), inhibitors of the enzyme aromatase,anti-androgens, and pharmaceutically acceptable salts, acids orderivatives of any of the above that act to regulate or inhibit hormoneaction on tumors.

Anti-Hormonal Agents

Examples of anti-estrogens and SERMs include, for example, tamoxifen(including NOLVADEX™), raloxifene, droloxifene, 4-hydroxytamoxifen,trioxifene, keoxifene, LY117018, onapristone, and toremifene(FARESTON®).

Inhibitors of the enzyme aromatase regulate estrogen production in theadrenal glands. Examples include 4(5)-imidazoles, aminoglutethimide,megestrol acetate (MEGACE®), exemestane, formestane, fadrozole, vorozole(RIVISOR®), letrozole (FEMARA®), and anastrozole (ARIMIDEX®).

Examples of anti-androgens include apalutamide, abiraterone,enzalutamide, flutamide, galeterone, nilutamide, bicalutamide,leuprolide, goserelin, ODM-201, APC-100, ODM-204.

Examples of progesterone receptor antagonist include onapristone.

Anti-Angiogenic Agents

Anti-angiogenic agents include, but are not limited to, retinoid acidand derivatives thereof, 2-methoxyestradiol, ANGIOSTATIN®, ENDOSTATIN®,regorafenib, necuparanib, suramin, squalamine, tissue inhibitor ofmetalloproteinase-1, tissue inhibitor of metalloproteinase-2,plasminogen activator inhibitor-1, plasminogen activator inbibitor-2,cartilage-derived inhibitor, paclitaxel (nab-paclitaxel), plateletfactor 4, protamine sulphate (clupeine), sulphated chitin derivatives(prepared from queen crab shells), sulphated polysaccharidepeptidoglycan complex (sp-pg), staurosporine, modulators of matrixmetabolism including proline analogs such as 1-azetidine-2-carboxylicacid (LACA), cishydroxyproline, d,I-3,4-dehydroproline, thiaproline,α,α′-dipyridyl, beta-aminopropionitrile fumarate,4-propyl-5-(4-pyridinyl)-2(3h)-oxazolone, methotrexate, mitoxantrone,heparin, interferons, 2 macroglobulin-serum, chicken inhibitor ofmetalloproteinase-3 (ChIMP-3), chymostatin, beta-cyclodextrintetradecasulfate, eponemycin, fumagillin, gold sodium thiomalate,d-penicillamine, beta-1-anticollagenase-serum, alpha-2-antiplasmin,bisantrene, lobenzarit disodium, n-2-carboxyphenyl-4-chloroanthronilicacid disodium or “CCA”, thalidomide, angiostatic steroid, carboxyaminoimidazole, metalloproteinase inhibitors such as BB-94, inhibitorsof S100A9 such as tasquinimod. Other anti-angiogenesis agents includeantibodies, preferably monoclonal antibodies against these angiogenicgrowth factors: beta-FGF, alpha-FGF, FGF-5, VEGF isoforms, VEGF-C,HGF/SF, and Ang-1/Ang-2.

Anti-Fibrotic Agents

Anti-fibrotic agents include, but are not limited to, the compounds suchas beta-aminoproprionitrile (BAPN), as well as the compounds disclosedin U.S. Pat. No. 4,965,288 relating to inhibitors of lysyl oxidase andtheir use in the treatment of diseases and conditions associated withthe abnormal deposition of collagen and U.S. Pat. No. 4,997,854 relatingto compounds which inhibit LOX for the treatment of various pathologicalfibrotic states, which are herein incorporated by reference. Furtherexemplary inhibitors are described in U.S. Pat. No. 4,943,593 relatingto compounds such as 2-isobutyl-3-fluoro-, chloro-, or bromo-allylamine,U.S. Pat. Nos. 5,021,456, 5,059,714, 5,120,764, 5,182,297, 5,252,608relating to 2-(1-naphthyloxymemyl)-3-fluoroallylamine, and US2004-0248871, which are herein incorporated by reference.

Exemplary anti-fibrotic agents also include the primary amines reactingwith the carbonyl group of the active site of the lysyl oxidases, andmore particularly those which produce, after binding with the carbonyl,a product stabilized by resonance, such as the following primary amines:emylenemamine, hydrazine, phenylhydrazine, and their derivatives;semicarbazide and urea derivatives; aminonitriles such as BAPN or2-nitroethylamine; unsaturated or saturated haloamines such as2-bromo-ethylamine, 2-chloroethylamine, 2-trifluoroethylamine,3-bromopropylamine, and p-halobenzylamines; and selenohomocysteinelactone.

Other anti-fibrotic agents are copper chelating agents penetrating ornot penetrating the cells. Exemplary compounds include indirectinhibitors which block the aldehyde derivatives originating from theoxidative deamination of the lysyl and hydroxylysyl residues by thelysyl oxidases. Examples include the thiolamines, particularlyD-penicillamine, and its analogs such as2-amino-5-mercapto-5-methylhexanoic acid,D-2-amino-3-methyl-3-((2-acetamidoethyl)dithio)butanoic acid,p-2-amino-3-methyl-3-((2-aminoethyl)dithio)butanoic acid,sodium-4-((p-1-dimethyl-2-amino-2-carboxyethyl)dithio)butane sulphurate,2-acetamidoethyl-2-acetamidoethanethiol sulphanate, andsodium-4-mercaptobutanesulphinate trihydrate.

Immunotherapeutic Agents

The immunotherapeutic agents include and are not limited to therapeuticantibodies suitable for treating subjects. Some examples of therapeuticantibodies include abagovomab, ABP-980, adecatumumab, afutuzumab,alemtuzumab, altumomab, amatuximab, anatumomab, arcitumomab,bavituximab, bectumomab, bevacizumab, bivatuzumab, blinatumomab,brentuximab, cantuzumab, catumaxomab, CC49, cetuximab, citatuzumab,cixutumumab, clivatuzumab, conatumumab, dacetuzumab, dalotuzumab,daratumumab, detumomab, dinutuximab, drozitumab, duligotumab,dusigitumab, ecromeximab, elotuzumab, emibetuzumab, ensituximab,ertumaxomab, etaracizumab, farletuzumab, ficlatuzumab, figitumumab,flanvotumab, futuximab, ganitumab, gemtuzumab, girentuximab,glembatumumab, ibritumomab, igovomab, imgatuzumab, indatuximab,inotuzumab, intetumumab, ipilimumab (YERVOY®, MDX-010, BMS-734016, andMDX-101), iratumumab, labetuzumab, lexatumumab, lintuzumab,lorvotuzumab, lucatumumab, mapatumumab, matuzumab, milatuzumab,minretumomab, mitumomab, mogamulizumab, moxetumomab, naptumomab,namatumab, necitumumab, nimotuzumab, nofetumomab, OBI-833, obinutuzumab,ocaratuzumab, ofatumumab, olaratumab, onartuzumab, oportuzumab,oregovomab, panitumumab, parsatuzumab, pasudotox, patritumab,pemtumomab, pertuzumab, pintumomab, pritumumab, racotumomab, radretumab,ramucirumab (Cyramza®), rilotumumab, rituximab, robatumumab,samalizumab, satumomab, sibrotuzumab, siltuximab, solitomab, simtuzumab,tacatuzumab, taplitumomab, tenatumomab, teprotumumab, tigatuzumab,tositumomab, trastuzumab, tucotuzumab, ublituximab, veltuzumab,vorsetuzumab, votumumab, zalutumumab, and 3F8. Rituximab can be used fortreating indolent B-cell cancers, including marginal-zone lymphoma, WM,CLL and small lymphocytic lymphoma. A combination of Rituximab andchemotherapy agents is especially effective.

The exemplified therapeutic antibodies may be further labeled orcombined with a radioisotope particle such as indium-111, yttrium-90(90Y-clivatuzumab), or iodine-131.

Cancer Gene Therapy and Cell Therapy

Cancer Gene Therapy and Cell Therapy includes the insertion of a normalgene into cancer cells to replace a mutated or altered gene; geneticmodification to silence a mutated gene; genetic approaches to directlykill the cancer cells; including the infusion of immune cells designedto replace most of the subject's own immune system to enhance the immuneresponse to cancer cells, or activate the subject's own immune system (Tcells or Natural Killer cells) to kill cancer cells, or find and killthe cancer cells; genetic approaches to modify cellular activity tofurther alter endogenous immune responsiveness against cancer.

Gene Editors

Examples of genome editing system include a CRISPR/Cas9 system, a zincfinger nuclease system, a TALEN system, a homing endonucleases system,and a meganuclease system.

CAR-T cell therapy and TCR-T cell therapy

CAR-T cell therapy includes a population of immune effector cellsengineered to express a chimeric antigen receptor (CAR), wherein the CARcomprises a tumor antigen-binding domain. The immune effector cell is aT cell or an NK cell. TCR-T cell therapy includes TCR-T cells that areengineered to target tumor derived peptides present on the surface oftumor cells. Cells can be autologous or allogeneic.

In some embodiments, the CAR comprises an antigen binding domain, atransmembrane domain, and an intracellular signalling domain.

In some embodiments, the intracellular domain comprises a primarysignaling domain, a costimulatory domain, or both of a primary signalingdomain and a costimulatory domain.

In some embodiments, the primary signaling domain comprises a functionalsignaling domain of one or more proteins selected from the groupconsisting of CD3 zeta, CD3 gamma, CD3 delta, CD3 epsilon, common FcRgamma (FCERIG), FcR beta (Fc Epsilon Rlb), CD79a, CD79b, Fcgamma RIIa,DAP10, and DAP12.

In some embodiments, the costimulatory domain comprises a functionaldomain of one or more proteins selected from the group consisting ofCD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocytefunction-associated antigen-1 (LFA-I), CD2, CD7, LIGHT, NKG2C, B7-H3, aligand that specifically binds with CD83, CDS, ICAM-1, GITR, BAFFR, HVEM(LIGHTR), SLAMF7, NKp80 (KLRF1), CD160, CD19, CD4, CD8alpha, CD8beta,IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4,CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD1 ld, ITGAE, CD103, ITGAL, CD 1 la,LFA-1, ITGAM, CD1 lb, ITGAX, CD1 lc, ITGB1, CD29, ITGB2, CD18, LFA-1,ITGB7, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84,CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100(SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3),BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, NKp44,NKp30, NKp46, and NKG2D.

In some embodiments, the transmembrane domain comprises a transmembranedomain of a protein selected from the group consisting of the alpha,beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4,CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137,CD154, KIRDS2, OX40, CD2, CD27, LFA-1 (CD1 la, CD18), ICOS (CD278),4-1BB (CD137), GITR, CD40, BAFFR, HVEM (LIGHTR), SLAMF7, NKp80 (KLRF1),CD160, CD19, IL2R beta, IL2R gamma, IL7R u, ITGA1, VLA1, CD49a, ITGA4,IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD1 ld, ITGAE, CD103, ITGAL, CD1la, LFA-1, ITGAM, CD1 lb, ITGAX, CD1 lc, ITGB1, CD29, ITGB2, CD18,LFA-1, ITGB7, TNFR2, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96(Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100(SEMA4D), SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME(SLAMF8), SELPLG (CD162), LTBR, PAG/Cbp, NKp44, NKp30, NKp46, NKG2D, andNKG2C.

In some embodiments, the antigen binding domain binds a tumor antigen.

In some embodiments, the tumor antigen is selected from the groupconsisting of: CD19; CD123; CD22; CD30; CD171; CS-1 (also referred to asCD2 subset 1, CRACC, SLAMF7, CD319, and 19A24); C-type lectin-likemolecule-1 (CLL-1 or CLECLI); CD33; epidermal growth factor receptorvariant III (EGFRvIII); ganglioside G2 (GD2); ganglioside GD3 (aNeuSAc(2-8)aNeuSAc (2-3)bDGaip(1-4)bDGIcp(1-1)Cer); TNF receptor family memberB cell maturation (BCMA); Tn antigen ((Tn Ag) or (GaINAcu-Ser/Thr));prostate-specific membrane antigen (PSMA); Receptor tyrosine kinase-likeorphan receptor 1 (RORI); Fms-Like, Tyrosine Kinase 3 (FLT3);Tumor-associated glycoprotein 72 (TAG72); CD38; CD44v6; Carcinoembryonicantigen (CEA); Epithelial cell adhesion molecule (EPCAM); B7H3 (CD276);KIT (CD117); Interleukin-13 receptor subunit alpha-2 (IL-13Ra2 orCD213A2); Mesothelin; Interleukin 11 receptor alpha (IL-11Ra); prostatestem cell antigen (PSCA); Protease Serine 21 (Testisin or PRSS21);vascular endothelial growth factor receptor 2 (VEGFR2); Lewis(Y)antigen; CD24; Platelet-derived growth factor receptor beta(PDGFR-beta); Stage-specificembryonic antigen-4 (SSEA-4); CD20; deltalike 3 (DLL3); Folate receptor alpha; Receptor tyrosine-protein kinase,ERBB2 (Her2/neu); Mucin 1, cell surface associated (MUC1); epidermalgrowth factor receptor (EGFR); neural cell adhesion molecule (NCAM);Prostase; prostatic acid phosphatase (PAP); elongation factor 2 mutated(ELF2M); Ephrin B2; fibroblast activation protein alpha (FAP);insulin-like growth factor 1 receptor (IGF-I receptor), carbonicanhydrase IX (CAIX); Proteasome (Prosome, Macropain) Subunit, Beta Type,9 (LMP2); glycoprotein 100 (gp100); oncogene fusion protein consistingof breakpoint cluster region (BCR) and Abelson murineleukemia viraloncogene homolog 1 (Abl) (bcr-abl); tyrosinase; ephrin type-A receptor 2(EphA2); Fucosyl GM1; sialyl Lewis adhesion molecule (sLe); gangliosideGM3 (aNeuSAc(2-3)bDGalp(1-4)bDGlcp(1-1)Cer); transglutaminase 5 (TGS5);high molecular weight-melanomaassociatedantigen (HMWMAA); o-acetyl-GD2ganglioside (OAcGD2); Folate receptor beta; tumor endothelial marker 1(TEM1/CD248); tumor endothelial marker 7-related (TEM7R); sixtransmembrane epithelial antigen of the prostate I (STEAP1); claudin 6(CLDN6); thyroid stimulating hormone receptor (TSHR); G protein-coupledreceptor class C group 5, member D (GPRCSD); chromosome X open readingframe 61 (CXORF61); CD97; CD179a; anaplastic lymphoma kinase (ALK);Polysialic acid; placenta-specific 1 (PLAC1); hexasaccharide portion ofgloboH glycoceramide (GloboH); mammary gland differentiation antigen(NY-BR-1); uroplakin 2 (UPK2); Hepatitis A virus cellular receptor 1(HAVCR1); adrenoceptor beta 3 (ADRB3); pannexin 3 (PANX3); Gprotein-coupled receptor 20 (GPR20); lymphocyte antigen 6 complex, locusK 9 (LY6K); Olfactory receptor 51E2 (ORS IE2); TCR Gamma AlternateReading Frame Protein (TARP); Wilms tumor protein (WT1); Cancer/testisantigen 1 (NY-ESO-1); Cancer/testis antigen 2 (LAGE-la);Melanomaassociated antigen 1 (MAGE-A1); ETS translocation-variant gene6, located on chromosome 12p (ETV6-AML); sperm protein 17 (SPA17); XAntigen Family, Member 1A (XAGE1); angiopoietin-binding cell surfacereceptor 2 (Tie 2); melanoma cancer testis antigen-1 (MADCT-1); melanomacancer testis antigen-2 (MAD-CT-2); Fos-related antigen 1; tumor proteinp53, (p53); p53 mutant; prostein; survivin; telomerase; prostatecarcinoma tumor antigen-1 (PCTA-1 or Galectin 8), melanoma antigenrecognized by T cells 1 (MelanA or MARTI); Rat sarcoma (Ras) mutant;human Telomerase reverse transcriptase (hTERT); sarcoma translocationbreakpoints; melanoma inhibitor of apoptosis (ML-IAP); ERG(transmembrane protease, serine 2 (TMPRSS2) ETS fusion gene); N-Acetylglucosaminyl-transferase V (NA17); paired box protein Pax-3 (PAX3);Androgen receptor; Cyclin B1; v-myc avian myelocytomatosis viraloncogene neuroblastoma derived homolog (MYCN); Ras Homolog Family MemberC (RhoC); Tyrosinase-related protein 2 (TRP-2); Cytochrome P450 1B1 (CYPIBI); CCCTC-Binding Factor (Zinc Finger Protein)-Like (BORIS or Brotherof the Regulator of Imprinted Sites), Squamous Cell Carcinoma AntigenRecognized By T Cells 3 (SART3); Paired box protein Pax-5 (PAX5);proacrosin binding protein sp32 (OY-TES I); lymphocyte-specific proteintyrosine kinase (LCK); A kinase anchor protein 4 (AKAP-4); synovialsarcoma, X breakpoint 2 (SSX2); Receptor for Advanced GlycationEndproducts (RAGE-I); renal ubiquitous 1 (RUI); renal ubiquitous 2(RU2); legumain; human papilloma virus E6 (HPV E6); human papillomavirus E7 (HPV E7); intestinal carboxyl esterase; heat shock protein 70-2mutated (mut hsp70-2); CD79a; CD79b; CD72; Leukocyte-associatedimmunoglobulin-like receptor 1 (LAIRI); Fc fragment of IgA receptor(FCAR or CD89); Leukocyte immunoglobulin-like receptor subfamily Amember 2 (LILRA2); CD300 molecule-like family member f (CD300LF); C-typelectin domain family 12 member A (CLEC12A); bone marrow stromal cellantigen 2 (BST2); EGF-like module containing mucin-like hormonereceptor-like 2 (EMR2); lymphocyte antigen 75 (LY75); Glypican-3 (GPC3);Fc receptor-like 5 (FCRL5); and immunoglobulin lambda-like polypeptide 1(IGLL1).

In some embodiments, the tumor antigen is selected from CD150, 5T4,ActRIIA, B7, BMCA, CA-125, CCNA1, CD123, CD126, CD138, CD14, CD148,CD15, CD19, CD20, CD200, CD21, CD22, CD23, CD24, CD25, CD26, CD261,CD262, CD30, CD33, CD362, CD37, CD38, CD4, CD40, CD40L, CD44, CD46, CD5,CD52, CD53, CD54, CD56, CD66a-d, CD74, CD8, CD80, CD92, CE7, CS-1,CSPG4, ED-B fibronectin, EGFR, EGFRvIII, EGP-2, EGP-4, EPHa2, ErbB2,ErbB3, ErbB4, FBP, GD2, GD3, HER1-HER2 in combination, HER2-HER3 incombination, HERV-K, HIV-1 envelope glycoprotein gp120, HIV-1 envelopeglycoprotein gp41, HLA-DR, HM1.24, HMW-MAA, Her2, Her2/neu, IGF-1R,IL-11Ralpha, IL-13R-alpha2, IL-2, IL-22R-alpha, IL-6, IL-6R, Ia, Ii,L1-CAM, L1-cell adhesion molecule, Lewis Y, L1-CAM, MAGE A3, MAGE-A1,MART-1, MUC1, NKG2C ligands, NKG2D Ligands, NYESO-1, OEPHa2, PIGF, PSCA,PSMA, ROR1, T101, TAC, TAG72, TIM-3, TRAIL-R1, TRAIL-R1 (DR4), TRAIL-R2(DR5), VEGF, VEGFR2, WT-I, a G-protein coupled receptor,alphafetoprotein (AFP), an angiogenesis factor, an exogenous cognatebinding molecule (ExoCBM), oncogene product, anti-folate receptor,c-Met, carcinoembryonic antigen (CEA), cyclin (D 1), ephrinB2,epithelial tumor antigen, estrogen receptor, fetal acethycholine ereceptor, folate binding protein, gpl00, hepatitis B surface antigen,kappa chain, kappa light chain, kdr, lambda chain, livin,melanoma-associated antigen, mesothelin, mouse double minute 2 homolog(MDM2), mucin 16 (MUC16), mutated p53, mutated ras, necrosis antigens,oncofetal antigen, ROR2, progesterone receptor, prostate specificantigen, tEGFR, tenascin, P2-Microgiobuiin, Fc Receptor-like 5 (FcRL5).

Non limiting examples of cell therapies include Algenpantucel-L,Sipuleucel-T, (BPX-501) rivogenlecleucel U.S. Pat. No. 9,089,520,WO2016100236, AU-105, ACTR-087, activated allogeneic natural killercells CNDO-109-AANK, MG-4101, AU-101, BPX-601, FATE-NK100, LFU-835hematopoietic stem cells, Imilecleucel-T, baltaleucel-T, PNK-007,UCARTCS1, ET-1504, ET-1501, ET-1502, ET-190, CD19-ARTEMIS, ProHema,FT-1050-treated bone marrow stem cell therapy, CD4CARNK-92 cells,CryoStim, AlloStim, lentiviral transduced huCART-meso cells, CART-22cells, EGFRt/19-28z/4-1BBL CAR T cells, autologous 4H11-28z/fIL-12/EFGRtT cell, CCR5-SBC-728-HSPC, CAR4-1BBZ, CH-296, dnTGFbRII-NY-ESOc259T,Ad-RTS-IL-12, IMA-101, IMA-201, CARMA-0508, TT-18, CMD-501, CMD-503,CMD-504, CMD-502, CMD-601, CMD-602, CSG-005.

In some embodiments, the tumor targeting antigen includes:Alpha-fetoprotein, such as ET-1402, and AFP-TCR; Anthrax toxin receptor1, such as anti-TEM8 CAR T-cell therapy; B cell maturation antigens(BCMA), such as bb-2121, UCART-BCMA, ET-140, KITE-585, MCM-998,LCAR-B38M, CART-BCMA, SEA-BCMA, BB212, UCART-BCMA, ET-140, P-BCMA-101,AUTO-2 (APRIL-CAR); Anti-CLL-1 antibodies, such as KITE-796; B7 homolog6, such as CAR-NKp30 and CAR-B7H6; B-lymphocyte antigen CD19, such asTBI-1501, CTL-119 huCART-19 T cells, JCAR-015 U.S. Pat. No. 7,446,190,JCAR-014, JCAR-017, (WO2016196388, WO2016033570, WO2015157386),axicabtagene ciloleucel (KTE-C19), U.S. Pat. Nos. 7,741,465, 6,319,494,UCART-19, EBV-CTL, T tisagenlecleucel-T (CTL019), WO2012079000,WO2017049166, CD19CAR-CD28-CD3zeta-EGFRt-expressing T cells, CD19/4-1BBLarmored CAR T cell therapy, C-CAR-011, CIK-CAR.CD19, CD19CAR-28-zeta Tcells, PCAR-019, MatchCART, DSCAR-01, IM19 CAR-T; B-lymphocyte antigenCD20, such as ATTCK-20; B-lymphocyte cell adhesion, such as UCART-22,JCAR-018 WO2016090190; NY-ESO-1, such as GSK-3377794, TBI-1301; Carbonicanhydrase, such as DC-Ad-GMCAIX; Caspase 9 suicide gene, such asCaspaCIDe DLI, BPX-501; CCR5, such as SB-728; CDw123, such as MB-102,UCART-123; CD20m such as CBM-C20.1; CD4, such as ICG-122; CD30, such asCART30 (CBM-C30.1; CD33, such as CIK-CAR.CD33; CD38, such as T-007,UCART-38; CD40 ligand, such as BPX-201; CEACAM protein 4 modulators,such as MG7-CART; Claudin 6, such as CSG-002; EBV targeted, such asCMD-003; EGFR, such as autologous 4H11-28z/fIL-12/EFGRt T cell;Endonuclease, such as PGN-514, PGN-201; Epstein-Barr virus specificT-lymphocytes, such as TT-10; Erbb2, such as CST-102, CIDeCAR;Ganglioside (GD2), such as 4SCAR-GD2; Glutamate carboxypeptidase II,such as CIK-CAR. PSMA, CART-PSMA-TGFβRDN, P-PSMA-101; Glypican-3 (GPC3),such as TT-16, GLYCAR; Hemoglobin, such as PGN-236; Hepatocyte growthfactor receptor, such as anti-cMet RNA CAR T; Human papillomavirus E7protein, such as KITE-439; Immunoglobulin gamma Fc receptor III, such asACTR087; IL-12, such as DC-RTS-IL-12; IL-12 agonist/mucin 16, such asJCAR-020; IL-13 alpha 2, such as MB-101; IL-2, such as CST-101; K-RasGTPase, such as anti-KRAS G12V mTCR cell therapy; Neural cell adhesionmolecule L1 L1CAM (CD171), such as JCAR-023; Latent membrane protein1/Latent membrane protein 2, such as Ad5f35-LMPd1-2-transducedautologous dendritic cells; Melanoma associated antigen 10, such asMAGE-A10C796T MAGE-A10 TCR; Melanoma associated antigen 3/Melanomaassociated antigen 6 (MAGE A3/A6) such as KITE-718; Mesothelin, such asCSG-MESO, TC-210; NKG2D, such as NKR-2; Ntrkr1 tyrosine kinase receptor,such as JCAR-024; T cell receptors, such as BPX-701, IMCgp100;T-lymphocyte, such as TT-12; Tumor infiltrating lymphocytes, such asLN-144, LN-145; and Wilms tumor protein, such as JTCR-016, WT1-CTL.

Lymphoma or Leukemia Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating lymphoma or leukemia. These agents include aldesleukin,alvocidib, amifostine trihydrate, aminocamptothecin, antineoplaston A10,antineoplaston AS2-1, anti-thymocyte globulin, arsenic trioxide, Bcl-2family protein inhibitor ABT-263, beta alethine, BMS-345541, bortezomib(VELCADE®), bortezomib (VELCADE®, PS-341), bryostatin 1, bulsulfan,campath-1H, carboplatin, carfilzomib (Kyprolis®), carmustine,caspofungin acetate, CC-5103, chlorambucil, CHOP (cyclophosphamide,doxorubicin, vincristine, and prednisone), cisplatin, cladribine,clofarabine, curcumin, CVP (cyclophosphamide, vincristine, andprednisone), cyclophosphamide, cyclosporine, cytarabine, denileukindiftitox, dexamethasone, docetaxel, dolastatin 10, doxorubicin,doxorubicin hydrochloride, DT-PACE (dexamethasone, thalidomide,cisplatin, doxorubicin, cyclophosphamide, and etoposide), enzastaurin,epoetin alfa, etoposide, everolimus (RAD001), FCM (fludarabine,cyclophosphamide, and mitoxantrone), FCR (fludarabine, cyclophosphamide,and rituximab), fenretinide, filgrastim, flavopiridol, fludarabine, FR(fludarabine and rituximab), geldanamycin (17-AAG), hyperCVAD(hyperfractionated cyclophosphamide, vincristine, doxorubicin,dexamethasone, methotrexate, and cytarabine), ICE (iphosphamide,carboplatin, and etoposide), ifosfamide, irinotecan hydrochloride,interferon alpha-2b, ixabepilone, lenalidomide (REVLIMID®, CC-5013),lymphokine-activated killer cells, MCP (mitoxantrone, chlorambucil, andprednisolone), melphalan, mesna, methotrexate, mitoxantronehydrochloride, motexafin gadolinium, mycophenolate mofetil, nelarabine,obatoclax (GX15-070), oblimersen, octreotide acetate, omega-3 fattyacids, Omr-IgG-am (WNIG, Omrix), oxaliplatin, paclitaxel, palbociclib(PD0332991), pegfilgrastim, PEGylated liposomal doxorubicinhydrochloride, perifosin, prednisolone, prednisone, recombinant flt3ligand, recombinant human thrombopoietin, recombinant interferon alfa,recombinant interleukin-11, recombinant interleukin-12, rituximab,R-CHOP (rituximab and CHOP), R-CVP (rituximab and CVP), R-FCM (rituximaband FCM), R-ICE (rituximab and ICE), and R-MCP (rituximab and MCP),R-roscovitine (seliciclib, CYC202), sargramostim, sildenafil citrate,simvastatin, sirolimus, styryl sulphones, tacrolimus, tanespimycin,temsirolimus (CC1-779), thalidomide, therapeutic allogeneic lymphocytes,thiotepa, tipifamib, vincristine, vincristine sulfate, vinorelbineditartrate, SAHA (suberanilohydroxamic acid, or suberoyl, anilide, andhydroxamic acid), vemurafenib (Zelboraf®), venetoclax (ABT-199).

One modified approach is radioimmunotherapy, wherein a monoclonalantibody is combined with a radioisotope particle, such as indium-111,yttrium-90, and iodine-131. Examples of combination therapies include,but are not limited to, iodine-131 tositumomab (BEXXAR®), yttrium-90ibritumomab tiuxetan (ZEVALIN®), and BEXXAR® with CHOP.

The abovementioned therapies can be supplemented or combined with stemcell transplantation or treatment. Therapeutic procedures includeperipheral blood stem cell transplantation, autologous hematopoieticstem cell transplantation, autologous bone marrow transplantation,antibody therapy, biological therapy, enzyme inhibitor therapy, totalbody irradiation, infusion of stem cells, bone marrow ablation with stemcell support, in vitro-treated peripheral blood stem celltransplantation, umbilical cord blood transplantation, immunoenzymetechnique, low-LET cobalt-60 gamma ray therapy, bleomycin, conventionalsurgery, radiation therapy, and nonmyeloablative allogeneichematopoietic stem cell transplantation.

Non-Hodgkin's Lymphomas Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating non-Hodgkin's lymphomas (NHL), especially those of B cellorigin, which include monoclonal antibodies, standard chemotherapyapproaches (e.g., CHOP, CVP, FCM, MCP, and the like),radioimmunotherapy, and combinations thereof, especially integration ofan antibody therapy with chemotherapy.

Examples of unconjugated monoclonal antibodies for the treatment ofNHL/B-cell cancers include rituximab, alemtuzumab, human or humanizedanti-CD20 antibodies, lumiliximab, anti-TNF-related apoptosis-inducingligand (anti-TRAIL), bevacizumab, galiximab, epratuzumab, SGN-40, andanti-CD74.

Examples of experimental antibody agents used in treatment of NHL/B-cellcancers include ofatumumab, ha20, PRO131921, alemtuzumab, galiximab,SGN-40, CHIR-12.12, epratuzumab, lumiliximab, apolizumab, milatuzumab,and bevacizumab.

Examples of standard regimens of chemotherapy for NHL/B-cell cancersinclude CHOP, FCM, CVP, MCP, R-CHOP, R-FCM, R-CVP, and R-MCP.

Examples of radioimmunotherapy for NHL/B-cell cancers include yttrium-90ibritumomab tiuxetan (ZEVALIN®) and iodine-131 tositumomab (BEXXAR®).

Mantle Cell Lymphoma Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating mantle cell lymphoma (MCL), which include combinationchemotherapies such as CHOP, hyperCVAD, and FCM. These regimens can alsobe supplemented with the monoclonal antibody rituximab to formcombination therapies R-CHOP, hyperCVAD-R, and R-FCM. Any of theabovementioned therapies may be combined with stem cell transplantationor ICE in order to treat MCL.

Other examples of therapeutic agents suitable for treating MCL include:

-   -   immunotherapy, such as monoclonal antibodies (like rituximab)        and cancer vaccines, such as GTOP-99, which are based on the        genetic makeup of an individual subject's tumor;    -   radioimmunotherapy, wherein a monoclonal antibody is combined        with a radioisotope particle, such as iodine-131 tositumomab        (BEXXAR®), yttrium-90 ibritumomab tiuxetan (ZEVALIN®), and        BEXXAR® in sequential treatment with CHOP;    -   autologous stem cell transplantation coupled with high-dose        chemotherapy, administering proteasome inhibitors such as        bortezomib (VELCADE® or PS-341), or administering        antiangiogenesis agents such as thalidomide, especially in        combination with rituximab;    -   drugs that lead to the degradation of Bcl-2 protein and increase        cancer cell sensitivity to chemotherapy, such as oblimersen, in        combination with other chemotherapeutic agents;    -   mTOR inhibitors, which can lead to inhibition of cell growth and        even cell death. Non-limiting examples are sirolimus,        temsirolimus (TORISEL®, CCI-779), CC-115, CC-223, SF-1126,        PQR-309 (bimiralisib), voxtalisib, GSK-2126458, and temsirolimus        in combination with RITUXAN®, VELCADE®, or other        chemotherapeutic agents;    -   other agents such as flavopiridol, palbociclib (PD0332991),        R-roscovitine (selicicilib, CYC202), styryl sulphones, obatoclax        (GX15-070), TRAIL, Anti-TRAIL death receptors DR4 and DR5        antibodies, temsirolimus (TORISEL®, CC1-779), everolimus        (RAD001), BMS-345541, curcumin, SAHA, thalidomide, lenalidomide        (REVLIMID®, CC-5013), and geldanamycin (17-AAG).

Waldenstrom's Macroglobulinemia Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating Waldenstrom's Macroglobulinemia (WM), which includealdesleukin, alemtuzumab, alvocidib, amifostine trihydrate,aminocamptothecin, antineoplaston A10, antineoplaston AS2-1,anti-thymocyte globulin, arsenic trioxide, autologous humantumor-derived HSPPC-96, Bcl-2 family protein inhibitor ABT-263, betaalethine, bortezomib (VELCADE®), bryostatin 1, busulfan, campath-1H,carboplatin, carmustine, caspofungin acetate, CC-5103, cisplatin,clofarabine, cyclophosphamide, cyclosporine, cytarabine, denileukindiftitox, dexamethasone, docetaxel, dolastatin 10, doxorubicinhydrochloride, DT-PACE, enzastaurin, epoetin alfa, epratuzumab(hLL2-anti-CD22 humanized antibody), etoposide, everolimus, fenretinide,filgrastim, fludarabine, ifosfamide, indium-111 monoclonal antibodyMN-14, iodine-131 tositumomab, irinotecan hydrochloride, ixabepilone,lymphokine-activated killer cells, melphalan, mesna, methotrexate,mitoxantrone hydrochloride, monoclonal antibody CD19 (such astisagenlecleucel-T, CART-19, CTL-019), monoclonal antibody CD20,motexafin gadolinium, mycophenolate mofetil, nelarabine, oblimersen,octreotide acetate, omega-3 fatty acids, oxaliplatin, paclitaxel,pegfilgrastim, PEGylated liposomal doxorubicin hydrochloride,pentostatin, perifosine, prednisone, recombinant flt3 ligand,recombinant human thrombopoietin, recombinant interferon alfa,recombinant interleukin-11, recombinant interleukin-12, rituximab,sargramostim, sildenafil citrate (VIAGRA®), simvastatin, sirolimus,tacrolimus, tanespimycin, thalidomide, therapeutic allogeneiclymphocytes, thiotepa, tipifamib, tositumomab, veltuzumab, vincristinesulfate, vinorelbine ditartrate, vorinostat, WT1 126-134 peptidevaccine, WT-1 analog peptide vaccine, yttrium-90 ibritumomab tiuxetan,yttrium-90 humanized epratuzumab, and any combination thereof.

Other examples of therapeutic procedures used to treat WM includeperipheral blood stem cell transplantation, autologous hematopoieticstem cell transplantation, autologous bone marrow transplantation,antibody therapy, biological therapy, enzyme inhibitor therapy, totalbody irradiation, infusion of stem cells, bone marrow ablation with stemcell support, in vitro-treated peripheral blood stem celltransplantation, umbilical cord blood transplantation, immunoenzymetechniques, low-LET cobalt-60 gamma ray therapy, bleomycin, conventionalsurgery, radiation therapy, and nonmyeloablative allogeneichematopoietic stem cell transplantation.

Diffuse Large B-Cell Lymphoma Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating diffuse large B-cell lymphoma (DLBCL), which includecyclophosphamide, doxorubicin, vincristine, prednisone, anti-CD20monoclonal antibodies, etoposide, bleomycin, many of the agents listedfor WM, and any combination thereof, such as ICE and R-ICE.

Chronic Lymphocytic Leukemia Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating chronic lymphocytic leukemia (CLL), which include chlorambucil,cyclophosphamide, fludarabine, pentostatin, cladribine, doxorubicin,vincristine, prednisone, prednisolone, alemtuzumab, many of the agentslisted for WM, and combination chemotherapy and chemoimmunotherapy,including the following common combination regimens: CVP, R-CVP, ICE,R-ICE, FCR, and FR.

Myelofibrosis Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating myelofibrosis, which include hedgehog inhibitors, histonedeacetylase (HDAC) inhibitors, and tyrosine kinase inhibitors.Non-limiting examples of hedgehog inhibitors are saridegib andvismodegib.

Examples of HDAC inhibitors include, but are not limited to, pracinostatand panobinostat.

Non-limiting examples of tyrosine kinase inhibitors are lestaurtinib,bosutinib, imatinib, gilteritinib, radotinib, and cabozantinib.

Hyperproliferative Disease Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating a hyperproliferative disease, which include gemcitabine,nab-paclitaxel, and gemcitabine/nab-paclitaxel with a JAK inhibitorand/or PI3Kδ inhibitor.

Bladder Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating bladder cancer, which include atezolizumab, carboplatin,cisplatin, docetaxel, doxorubicin, fluorouracil (5-FU), gemcitabine,idosfamide, Interferon alfa-2b, methotrexate, mitomycin, nab-paclitaxel,paclitaxel, pemetrexed, thiotepa, vinblastine, and any combinationthereof.

Breast Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating breast cancer, which include albumin-bound paclitaxel,anastrozole, capecitabine, carboplatin, cisplatin, cyclophosphamide,docetaxel, doxorubicin, epirubicin, everolimus, exemestane,fluorouracil, fulvestrant, gemcitabine, Ixabepilone, lapatinib,Letrozole, methotrexate, mitoxantrone, paclitaxel, pegylated liposomaldoxorubicin, pertuzumab, tamoxifen, toremifene, trastuzumab,vinorelbine, and any combinations thereof.

Triple Negative Breast Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating triple negative breast cancer, which include cyclophosphamide,docetaxel, doxorubicin, epirubicin, fluorouracil, paclitaxel, andcombinations thereof.

Colorectal Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating colorectal cancer, which include bevacizumab, capecitabine,cetuximab, fluorouracil, irinotecan, leucovorin, oxaliplatin,panitumumab, ziv-aflibercept, and any combinations thereof.

Castration-Resistant Prostate Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating castration-resistant prostate cancer, which includeabiraterone, cabazitaxel, docetaxel, enzalutamide, prednisone,sipuleucel-T, and any combinations thereof.

Esophageal and Esophagogastric Junction Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating esophageal and esophagogastric junction cancer, which includecapecitabine, carboplatin, cisplatin, docetaxel, epirubicin,fluoropyrimidine, fluorouracil, irinotecan, leucovorin, oxaliplatin,paclitaxel, ramucirumab, trastuzumab, and any combinations thereof.

Gastric Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating gastric cancer, which include capecitabine, carboplatin,cisplatin, docetaxel, epirubicin, fluoropyrimidine, fluorouracil,Irinotecan, leucovorin, mitomycin, oxaliplatin, paclitaxel, ramucirumab,trastuzumab, and any combinations thereof.

Head & Neck Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating head & neck cancer, which include afatinib, bleomycin,capecitabine, carboplatin, cetuximab, cisplatin, docetaxel,fluorouracil, gemcitabine, hydroxyurea, methotrexate, nivolumab,paclitaxel, pembrolizumab, vinorelbine, and any combinations thereof.

Hepatobiliary Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating hepatobiliary cancer, which include capecitabine, cisplatin,fluoropyrimidine, 5-fluorourcil, gemecitabine, oxaliplatin, sorafenib,and any combinations thereof.

Hepatocellular Carcinoma Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating hepatocellular carcinoma, which include capecitabine,doxorubicin, gemcitabine, sorafenib, and any combinations thereof.

Non-Small Cell Lung Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating non-small cell lung cancer (NSCLC), which include afatinib,albumin-bound paclitaxel, alectinib, bevacizumab, bevacizumab,cabozantinib, carboplatin, cisplatin, crizotinib, dabrafenib, docetaxel,erlotinib, etoposide, gemcitabine, nivolumab, paclitaxel, pembrolizumab,pemetrexed, ramucirumab, trametinib, trastuzumab, vandetanib,vemurafenib, vinblastine, vinorelbine, and any combinations thereof.

Small Cell Lung Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating small cell lung cancer (SCLC), which include bendamustime,carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin,etoposide, gemcitabine, ipillimumab, irinotecan, nivolumab, paclitaxel,temozolomide, topotecan, vincristine, vinorelbine, and any combinationsthereof.

Melanoma Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating melanoma, which include albumin bound paclitaxel, carboplatin,cisplatin, cobiemtinib, dabrafenib, dacrabazine, IL-2, imatinib,interferon alfa-2b, ipilimumab, nitrosourea, nivolumab, paclitaxel,pembrolizumab, pilimumab, temozolomide, trametinib, vemurafenib,vinblastine, and any combinations thereof.

Ovarian Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating ovarian cancer, which include 5-flourouracil, albumin boundpaclitaxel, altretamine, anastrozole, bevacizumab, capecitabine,carboplatin, cisplatin, cyclophosphamide, docetaxel, doxorubicin,etoposide, exemestane, gemcibabine, ifosfamide, irinotecan, letrozole,leuprolide acetate, liposomal doxorubicin, megestrol acetate, melphalan,olaparib, oxaliplatin, paclitaxel, Pazopanib, pemetrexed, tamoxifen,topotecan, vinorelbine, and any combinations thereof.

Pancreatic Cancer Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating pancreatic cancer, which include 5-fluorourcil, albumin-boundpaclitaxel, capecitabine, cisplatin, docetaxel, erlotinib,fluoropyrimidine, gemcitabine, irinotecan, leucovorin, oxaliplatin,paclitaxel, and any combinations thereof.

Renal Cell Carcinoma Combination Therapy

In some embodiments, the additional therapeutic agents are suitable fortreating renal cell carcinoma, which include axitinib, bevacizumab,cabozantinib, erlotinib, everolimus, levantinib, nivolumab, pazopanib,sorafenib, sunitinib, temsirolimus, and any combinations thereof.

VIII. Kits

The present disclosure provides a kit comprising a compound of thepresent disclosure or a pharmaceutically acceptable salt thereof. Thekit may further comprise instructions for use, e.g., for use in treatinga viral infection. The instructions for use are generally writteninstructions, although electronic storage media (e.g., magnetic disketteor optical disk) containing instructions are also acceptable.

The present disclosure also provides a pharmaceutical kit comprising oneor more containers comprising a compound of the present disclosure or apharmaceutically acceptable salt thereof. Optionally associated withsuch container(s) can be a notice in the form prescribed by agovernmental agency regulating the manufacture, use or sale ofpharmaceuticals, which notice reflects approval by the agency for themanufacture, use or sale for human administration. Each component (ifthere is more than one component) can be packaged in separate containersor some components can be combined in one container wherecross-reactivity and shelf life permit. The kits may be in unit dosageforms, bulk packages (e.g., multi-dose packages) or sub-unit doses. Kitsmay also include multiple unit doses of the compounds and instructionsfor use and be packaged in quantities sufficient for storage and use inpharmacies (e.g., hospital pharmacies and compounding pharmacies).

Also provided are articles of manufacture comprising a unit dosage of acompound of the present disclosure or a pharmaceutically acceptable saltthereof, in suitable packaging for use in the methods described herein.Suitable packaging is known in the art and includes, for example, vials,vessels, ampules, bottles, jars, flexible packaging and the like. Anarticle of manufacture may further be sterilized and/or sealed.

IX. Examples

The embodiments are also directed to processes and intermediates usefulfor preparing the subject compounds or pharmaceutically acceptable saltsthereof.

Many general references providing commonly known chemical syntheticschemes and conditions useful for synthesizing the disclosed compoundsare available (see, e.g., Smith, March's Advanced Organic Chemistry:Reactions, Mechanisms, and Structure, 7^(th) edition,Wiley-Interscience, 2013.)

Compounds as described herein can be purified by any of the means knownin the art, including chromatographic means, such as high performanceliquid chromatography (HPLC), preparative thin layer chromatography,flash column chromatography and ion exchange chromatography. Anysuitable stationary phase can be used, including normal and reversedphases as well as ionic resins. Most typically the disclosed compoundsare purified via silica gel and/or alumina chromatography. See, e.g.,Introduction to Modern Liquid Chromatography, 2nd ed., ed. L. R. Snyderand J. J. Kirkland, John Wiley and Sons, 1979; and Thin LayerChromatography, E. Stahl (ed.), Springer-Verlag, New York, 1969.

Compounds were characterized using standard instrumentation methods. NMRspectra were measured on a Bruker AVANCE-600 instrument (¹H at 600.13MHz) with a cryoprobe and a Bruker AVANCE-500 instrument with acryoprobe (³¹P at 202.4 MHz, ¹⁹F at 470.4 MHz). Mass spectrometry wasobtained on a Waters Q-Tof Micro in electron spray ionization (ESI)mode. HPLC was obtained on Waters LC-MS instrument (Waters 600controller, Waters 3100 Mass detector, Waters Photodiodarray detector)on Luna C18 column (Phenomenex, 5 μm, 150×4.6 mm) and Zic-Hilic column(SeQuant, 5 μm, 100×4.6 mm).

During any of the processes for preparation of the subject compounds, itmay be necessary and/or desirable to protect sensitive or reactivegroups on any of the molecules concerned. This may be achieved by meansof conventional protecting groups as described in standard works, suchas T. W. Greene and P. G. M. Wuts, “Protective Groups in OrganicSynthesis,” 4^(th) ed., Wiley, New York 2006. The protecting groups maybe removed at a convenient subsequent stage using methods known from theart.

Exemplary chemical entities useful in methods of the embodiments willnow be described by reference to illustrative synthetic schemes fortheir general preparation herein and the specific examples that follow.Artisans will recognize that, to obtain the various compounds herein,starting materials may be suitably selected so that the ultimatelydesired substituents will be carried through the reaction scheme with orwithout protection as appropriate to yield the desired product.Alternatively, it may be necessary or desirable to employ, in the placeof the ultimately desired substituent, a suitable group that may becarried through the reaction scheme and replaced as appropriate with thedesired substituent.

Furthermore, one of skill in the art will recognize that thetransformations shown in the schemes below may be performed in any orderthat is compatible with the functionality of the particular pendantgroups. Each of the reactions depicted in the general schemes ispreferably run at a temperature from about 0° C. to the refluxtemperature of the organic solvent used.

The Examples provided herein describe the synthesis of compoundsdisclosed herein as well as intermediates used to prepare the compounds.It is to be understood that individual steps described herein may becombined. It is also to be understood that separate batches of acompound may be combined and then carried forth in the next syntheticstep.

In the following description of the Examples, specific embodiments aredescribed. These embodiments are described in sufficient detail toenable those skilled in the art to practice certain embodiments of thepresent disclosure. Other embodiments may be utilized and logical andother changes may be made without departing from the scope of thedisclosure. The following description is, therefore, not intended tolimit the scope of the present disclosure.

The methods of the present invention generally provide a specificenantiomer or diastereomer as the desired product, although thestereochemistry of the enantiomer or diastereomer was not determined inall cases. When the stereochemistry of the specific stereocenter in theenantiomer or diastereomer is not determined, the compound is drawnwithout showing any stereochemistry at that specific stereocenter eventhough the compound can be substantially enantiomerically ordisatereomerically pure.

Representative syntheses of compounds of the present disclosure aredescribed in schemes below, and the particular examples that follow.Specific examples were named using PerkinElmer ChemDraw Professionalversion 17.0.0.206 (121), PerkinElmer Informatics, Inc.

Example 1. General Synthetic Method

The specific 3′3′-cyclic dinucleotides detailed in the Examples weresynthesized according to the general synthetic method described below.

List of Abbreviations

ACN acetonitrile

BOMOM benzoyloxymethoxymethyl

BSA bovine serum albumin

Bz benzoyl

CPG controlled pore glass

CSO (−)-(8,8-dichlorocamphorylsulfonyl)-oxaziridine

DBU 1,8-diazabicyclo[5.4.0]undec-7-en

DCA dichloroacetic acid

DCM dichloromethane

DEA diethylamine

DMAM dimethylaminomethylene

DMF dimethylformamide

DMOCP 2-chloro-5,5-dimethyl-1,3,2-dioxaphosphorinane-2-oxide

DMSO dimethylsulfoxide

DMTr 4,4-dimethoxytrityl

EtOH ethanol

ESI electron spray ionization

ETT ethylthiotetrazole

FBS fetal bovine serum

HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

HPLC high performance liquid chromatography

HRMS high resolution mass spectrometry

iBu isobutyl

iPr isopropyl

LCAA long chain aminoalkyl

MeIm 1-methylimidazole

MeOH methanol

MOP 4-methoxy-1-oxido-2-pyridylmethyl

MOP-OH 4-methoxy-1-oxido-2-pyridylmethanol

MPNO 4-methoxypyridine-1-oxide

NMR nuclear magnetic resonance

Pic 4-methoxy-2-pyridylmethyl

Pic-OH 4-methoxy-2-pyridylmethanol

Py pyridine

TBDMS tert-butyldimethylsilyl

TBDMSCl tert-butyldimethylsilyl chloride

tBuOOH tert-butyl hydroperoxide

TEA triethylamine

TEAB triethylammonium bicarbonate

THF tetrahydrofuran

TIPSCl triisopropylbenzenesulfonyl chloride

Ts p-toluenesulfonyl

Tzol tetrazole

Preparation of Monomers Derived from nucleoside-3′-methylphosphonates

Phosphonate and phosphoramidite monomers were prepared according to Páv,O; Košiová, I.; Barvik, I.; Pohl, R.; Buděšínský, M.; Rosenberg, I.Synthesis of oligoribonucleotides with phosphonate-modified linkages.Org. Biomol. Chem. 2011, 9, 6120-6126 and Páv, O.; Panova, N.; Snãšel,J.; Zbomiková, E.; Rosenberg I. Activation of human RNase L by 2′- and5′-O-methylphosphonate-modified oligoadenylates. Bioorg. Med. Chem.Lett. 2012, 22, 181-185.

2′-Deoxy-2′-fluoro phosphonate and phosphoramidite monomers shown asIntermediates 1 through 7 in Scheme 1 below were also prepared usingabove mentioned protocols from commercially available precursors(Carbosynth).

Compound characterization was consistent with the literature values.Exemplary data for the adenine (A) Intermediate 4 and the guanine (G)Intermediate 7 are shown in the table below.

Intermediate Characterization Data 4 ³¹P NMR (C₆D₆) δ 154.71 (d, J = 6.6Hz), 153.69 (d, J = 10.0 Hz). 7 HRMS (M-H)⁻ for C₄₂H₄₄FN₇O₁₁P: calcd872.28205, found 872.28184; ¹H NMR (DMSO-d₆) δ 11.39 (br s, 1H), 8.64(br s, 1H), 8.11 (d, J = 7.2 Hz, 1H), 8.00 (s, 1H), 7.25 (m, 2H), 7.19(m, 2H), 7.14 (m, 1H), 7.13 (m, 2H), 7.12 (m, 2H), 7.10 (d, J = 3.6 Hz,1H), 6.93 (dd, J = 7.2, 3.6 Hz, 1H), 6.76 (m, 2H), 6.75 (m, 2H), 6.19(dd, J = 20.8, 1.5 Hz, 1H), 5.92 (ddd, J = 52.2, 4.2, 1.5 Hz, 1H), 4.89(ddd, J = 22.4, 8.3, 4.2 Hz, 1H), 4.83 (dd, J = 17.3, 8.2 Hz, 1H), 4.79(dd, J = 17.3, 8.2 Hz, 1H), 4.07 (dd, J = 8.3, 5.0, 2.5 Hz, 1H), 3.75(s, 3H), 3.69 (s, 6H), 3.63 (dd, J = 13.1, 7.9 Hz, 1H), 3.55 (dd, J =13.1, 8.1 Hz, 1H), 3.24 (dd, J = 10.7, 2.5 Hz, 1H), 3.21 (dd, J = 10.7,5.0 Hz, 1H), 3.12 (s, 3H), 3.01 (d, J = 0.6 Hz, 3H); ³¹P NMR (DMSO-d₆) δ12.71; ¹⁹F NMR (DMSO-d₆) δ −197.52.

Representative syntheses of intermediates are described below.

Preparation of Intermediate 5

0.45M Tetrazole in ACN (7.1 mL, 3.2 mmol) was added under argon to astirred solution of compound 3 (1.0 g, 1.6 mmol) and methylN,N,N′,N′-tetraisopropylphosphorodiamidite (1.4 mL, 4.8 mmol) in DCM (20mL). The reaction mixture was stirred under argon for 2 h at r.t. Thereaction mixture was diluted with DCM (300 mL) and extracted withsaturated solution of sodium bicarbonate (3×100 ml). The organic layerwas dried over anhydrous sodium sulfate and evaporated. The residue wasdissolved in toluene (5 mL) and added dropwise into hexane (300 mL). Theprecipitate was collected and freeze-dried from benzene to yield 1.0 g(78%): ³¹P NMR (C₆D₆) δ 153.38 (d, J=8.5 Hz), 152.88 (d, J=6.5 Hz).

Preparation of Intermediate 6

Sodium hydride (0.72 g, 18.0 mmol) was added under argon at 4° C. to astirred solution of Intermediate 2 (4 g, 6 mmol) in DMF (60 mL) (Scheme1). The reaction mixture was stirred for additional 60 min at 4° C.Diisopropyl tosyloxymethylphosphonate (3.1 g, 9.0 mmol) was added to thereaction mixture, which was stirred under argon for additional 16 h atr.t., whereupon glacial AcOH (1.0 mL, 18.0 mmol) in DMF (10 mL) wasadded dropwise at 4° C. to the reaction mixture. The subsequent mixturewas evaporated and the crude phosphonate was purified by chromatographyon silica gel (elution with gradient of 0-10% ethanol in chloroform) toyield 3.5 g (69%): HRMS (ESI) calcd for C₄₅H₄₉O₉N₅FNaP (M+Na)⁺876.31441, found 876.31425.

Bromotrimethylsilane (2.2 mL; 16.4 mmol) was added to a solution of theabove prepared diisopropyl phosphonate (3.5 g; 4.1 mmol) and2,6-lutidine (3.8 mL; 32.8 mmol) in ACN (45 mL). The reaction mixturewas stirred for 16 h at r.t. and evaporated. The residue was dissolvedin chloroform (0.5 L) and extracted with 0.2 M TEAB (3×100 ml). Theorganic layer was dried over anhydrous sodium sulfate, evaporated andcoevaporated with dioxane and pyridine. The crude nucleoside phosphonicacid was used without further purification.

DMOCP (3.8 g; 20.5 mmol) was added to a solution of the above preparednucleoside phosphonic acid, MOP-OH (1.9 g; 12.3 mmol) and MPNO (2.6 g;20.5 mmol) in pyridine (45 mL). The reaction mixture was stirred for 16h at r.t., quenched by the addition of 2M TEAB (20 mL) and evaporated.The residue was dissolved in chloroform (0.5 L) and extracted with 0.2 MTEAB (3×100 ml). The organic layer was dried over anhydrous sodiumsulfate, evaporated and coevaporated with dioxane to afford the crudediMOP phosphonate, which was used without further purification.

The above diMOP phosphonate was treated 6 h at r.t. with benzenethiol (6mL) and TEA (8.4 mL) in dioxane (45 mL). The reaction mixture wasdiluted with ethyl acetate and directly purified by chromatography onsilica gel (elution with gradient of 0-100% ethylacetate/ethanol/acetone/water 4:1:1:1 in ethyl acetate (SiO₂ bufferedwith TEA). The resulting Intermediate 6 was lyophilized from dioxane toyield 2.2 g (59%): HRMS (M-H)⁻ for C₄₆H₄₃FN₆O₁₁P: calcd 905.27115, found905.27039; ¹H NMR (DMSO-d₆) δ 11.25 (br s, 1H), 8.74 (s, 1H), 8.67 (s,1H), 8.12 (d, J=7.2 Hz, 1H), 8.04 (m, 2H), 7.64 (m, 1H), 7.55 (m, 2H),7.27 (m, 2H), 7.19 (m, 2H), 7.14 (m, 4H), 7.135 (m, 1H), 7.12 (d, J=3.6Hz, 1H), 6.94 (dd, J=7.2, 3.6 Hz, 1H), 6.77 (m, 4H), 6.43 (dd, J=18.7,1.6 Hz, 1H), 6.01 (ddd, J=52.0, 4.0, 1.6 Hz, 1H), 4.91 (ddd, J=22.2,8.1, 4.0 Hz, 1H), 4.87 (dd, J=17.2, 8.0 Hz, 1H), 4.83 (dd, J=17.2, 8.2Hz, 1H), 4.19 (ddd, J=8.1, 4.8, 2.5 Hz, 1H), 3.76 (s, 3H), 3.68 (s, 3H),3.67 (s, 3H), 3.62 (m, 2H), 3.29 (dd, J=10.9, 2.5 Hz, 1H), 3.22 (dd,J=10.9, 4.8 Hz, 1H); ³¹P NMR (DMSO-d₆) δ 12.53; ¹⁹F NMR (DMSO-d₆) δ−199.74.

Preparation of Intermediate 8

Sodium hydride (0.72 g, 18.0 mmol) was added under argon at 4° C. to astirred solution of Intermediate 2 (4 g, 6 mmol) in DMF (60 mL) (Scheme1). The reaction mixture was stirred for additional 60 min at 4° C.Diisopropyl tosyloxymethylphosphonate (3.1 g, 9.0 mmol) was added to thereaction mixture, which was stirred under argon for additional 16 h atr.t., whereupon glacial AcOH (1.0 mL, 18.0 mmol) in DMF (10 mL) wasadded dropwise at 4° C. to the reaction mixture. The subsequent mixturewas evaporated and the crude phosphonate was purified by chromatographyon silica gel (elution with gradient of 0-10% ethanol in chloroform) toyield 3.5 g (69%): HRMS (ESI) calcd for C₄₅H₄₉O₉N₅FNaP (M+Na)⁺876.31441, found 876.31425.

Bromotrimethylsilane (2.2 mL; 16.4 mmol) was added to a solution of theabove prepared diisopropyl phosphonate (3.5 g; 4.1 mmol) and2,6-lutidine (3.8 mL; 32.8 mmol) in ACN (45 mL). The reaction mixturewas stirred for 16 h at r.t. and evaporated. The residue was dissolvedin chloroform (0.5 L) and extracted with 0.2 M TEAB (3×100 ml). Theorganic layer was dried over anhydrous sodium sulfate, evaporated andcoevaporated with dioxane and pyridine. The crude nucleoside phosphonicacid was used without further purification.

DMOCP (3.8 g; 20.5 mmol) was added to a solution of nucleosidephosphonic acid, Pic-OH (1.7 g; 12.3 mmol) and MPNO (2.6 g; 20.5 mmol)in pyridine (45 mL). The reaction mixture was stirred for 16 h at r.t.,quenched by the addition of 2M TEAB (20 mL) and evaporated. The residuewas dissolved in chloroform (0.5 L) and extracted with 0.2 M TEAB (3×100ml). The organic layer was dried over anhydrous sodium sulfate,evaporated and coevaporated with dioxane. The crude diPic phosphonatewas then treated 6 h at r.t. with benzenethiol (6 mL) and TEA (8.4 mL)in dioxane (45 mL) The reaction mixture was diluted with ethyl acetateand directly purified by chromatography on silica gel (elution withgradient of 0-100% ethyl acetate/ethanol/acetone/water 4:1:1:1 in ethylacetate (SiO₂ buffered with TEA). Intermediate 8 was lyophilized fromdioxane to yield 2.4 g (66%): HRMS (ESI) calcd for C₄₆H₄₃FN₆O₁₀P (M-H)⁻889.27623, found 889.27583; ¹H NMR (DMSO-d₆) δ 11.25 (br s, 1H), 8.74(s, 1H), 8.67 (s, 1H), 8.26 (d, J=5.7 Hz, 1H), 8.04 (m, 2H), 7.64 (m,1H), 7.54 (m, 2H), 7.27 (m, 2H), 7.19 (m, 2H), 7.14 (m, 5H), 7.04 (d,J=2.6 Hz, 1H), 6.81 (dd, J=5.7, 2.6 Hz, 1H), 6.76 (m, 4H), 6.43 (dd,J=18.6, 1.6 Hz, 1H), 6.01 (ddd, J=51.8, 4.0, 1.6 Hz, 1H), 4.90 (ddd,J=22.3, 8.1, 4.0 Hz, 1H), 4.80 (dd, J=14.3, 7.5 Hz, 1H), 4.78 (dd,J=14.3, 7.5 Hz, 1H), 4.20 (ddd, J=8.1, 5.0, 2.5 Hz, 1H), 3.77 (s, 3H),3.67 (s, 6H), 3.64 (m, 2H), 3.31 (dd, J=11.0, 2.5 Hz, 1H), 3.22 (dd,J=11.0, 5.0 Hz, 1H); ³¹P NMR (DMSO-d₆) δ 12.48; ¹⁹F NMR (DMSO-d₆) δ−199.61.

Preparation of Intermediate 9

Intermediate 8 (1.3 g, 1.5 mmol) was loaded on silica gel column in DCM(10 mL). The column was washed with DCA (10 mL, 3% in DCM) and aged for15 min at r.t. The column was subsequently washed with a mixture of DCA(25 mL, 3% in DCM)/10% EtOH in CHCl₃ (25 mL), and then with 10% EtOH inCHCl₃ (100 mL). The crude Intermediate 9 was eluted from the column with50% MeOH in H₂O (100 mL), evaporated, purified by preparative HPLC(elution with gradient of 0-50% methanol in water) and freeze-dried fromwater to yield 0.54 g (63%): HRMS (ESI) calcd for C₂₅H₂₅FN₆O₈P (M-H)⁻587.14555, found 587.14561; ¹H NMR (DMSO-d₆) δ 11.23 (br s, 1H), 8.82(s, 1H), 8.76 (s, 1H), 8.26 (d, J=5.6 Hz, 1H), 8.05 (m, 2H), 7.64 (m,1H), 7.55 (m, 2H), 7.11 (d, J=2.6 Hz, 1H), 6.81 (dd, J=5.6, 2.6 Hz, 1H),6.36 (d, J=16.1 Hz, 1H), 5.55 (dd, J=53.1, 3.7 Hz, 1H), 4.82 (ddd,J=26.5, 9.2, 3.7 Hz, 1H), 4.82 (dd, J=14.5, 7.7 Hz, 1H), 4.80 (dd,J=14.5, 7.7 Hz, 1H), 4.00 (dt, J=9.2, 2.1 Hz, 1H), 3.82 (s, 3H), 3.82(dd, J=13.2, 2.1 Hz, 1H), 3.78 (dd, J=13.2, 2.1 Hz, 1H), 3.64 (dd,J=14.2, 8.0 Hz, 1H), 3.60 (dd, J=14.2, 3.5 Hz, 1H); ³¹P NMR (DMSO-d₆) δ13.65; ¹⁹F NMR (DMSO-d₆) δ −198.08.

Preparation of a Modified Solid Support CE-CPG

The solid support (LCAA-CPG) modified with12-cyano-13-[(4,4′-dimethoxytrityl)oxy]-3,6,9-trioxatridecanehydrogensuccinate (CE-CPG) was prepared according to Pačes, O., et al.(2008). Collection of Czechoslovak Chemical Communications 73(1): 32-43.

CE-CPG

Synthesis of Dinucleotides Derived from nucleoside-3′-methylphosphonates

The dinucleotides were synthesized by “trityl off” method in a 1 μmolscale in the 5′→2′ direction using the CE-CPG (20 mg), see Scheme 1. Thesynthetic protocols for phosphotriester and phosphoramidite methods areshown in Table 1. The capping step was optional in many instances. In atypical experiment, six columns were used in the preparation of a CDN.The synthesis was performed on MOS oligonucleotide synthesizer (IOCBPrague, Czech Republic). The average yield of the coupling step was inthe range 93-95% (conductivity detector, DMTr⁺).

TABLE 1 Protocol for the synthesis of dinucleotides Operation AgentVolume (ml) Time (s) Phosphotriester condensation method 1.Detritylation 3% CHCl₂COOH in DCM 3 135 2. Condensation 0.1 mol · l⁻¹monomer 0.1 600 in pyridine 0.3 mol · l⁻¹ TIPSCl 0.1 in pyridine 3.Capping Ac₂O/pyridine/THF 1:1:8 0.1 150 1-MeIm/THF 1:9 0.1Phosphoramidite condensation method 1. Detritylation 3% CHCl₂COOH in DCM3 135 2. Condensation 0.1 mol · l⁻¹ monomer 0.1 600 in ACN 0.3 mol · l⁻¹ETT in ACN 0.1 3. Capping Ac₂O/pyridine/THF 1:1:8 0.1 150 1-MeIm/THF 1:90.1 4. Oxidation tBuOOH/DCM 1:4 0.2 180Cyclization. Deprotection and Purification of Dinucleotides

10% solution of Et₂NH in ACN (500 μl) was added to the CE-CPG solidsupport with bound linear dimer, and the heterogeneous mixture wasshaken on an Eppendorf Thermomixer comfort shaker for 2 h at roomtemperature. The solvents were evaporated and a solution of DMOCP (10mg) in pyridine (400 μl) was added. The heterogeneous mixture was shakenfor 2 hours at room temperature. Then, 10 μl of Et₃N and 266 μl of waterwere added to the reaction mixture and the shaking was continued for 16hours at 65° C. The reaction mixture was evaporated.

50% Aqueous MeOH (600 μl) was added to the reaction mixture and thesolid support was separated from the solution by decantation and washedwith 50% aqueous MeOH (3×200 μl). A solution of 33% MeNH₂ in EtOH (600μl) was added to the methanolic solution and the mixture was shaken for8 hours at room temperature. The solvents were then evaporated. Theresidue was dissolved in 0.1 mol·l⁻¹ TEAB (1.5 ml) and applied to asemipreparative C18 column. Preparative HPLC conditions used forpurification were as follows: Luna C18 (5 μm, 250×10 mm, Phenomenex);flow 3 mL/min; mobile phase A: 0.1M TEAB/H₂O, mobile phase B: 50%ACN/0.1M TEAB/H₂O; HPLC method 1: isocratic elution A (15 min), thenlinear gradient A−30% B (35 min); HPLC method 2: isocratic elution A (15min), then linear gradient A−20% B (35 min).

Example 2.2-amino-9-((2R,3R,3aS,7aR,9R,10R,10aS,15aR)-2-(6-amino-9H-purin-9-yl)-3,5,10,13-tetrahydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecin-9-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared according to the solid phase method describedabove without capping steps.

ESI-MS (M-H)⁻ for C₂₁H₂₅N₁₀O₁₃P₂ calculated: 687.1; found: 687.2; HPLCMethod 1, retention time (min)=33.6.

Example 3.2-amino-9-((2R,3R,3aS,7aR,9R,10R,10aS,15aR)-9-(6-amino-9H-purin-9-yl)-3,5,10,13-tetrahydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecin-2-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared on solid phase according to the methoddescribed above without capping steps.

ESI-MS (M-H)⁻ for C₂₁H₂₅N₁₀O₁₃P₂ calculated: 687.1; found: 687.1; HPLCMethod 1, retention time (min)=32.3.

Example 4.2-amino-9-((2R,3R,3aR,7aR,9R,10R,10aS,15aR)-2-(6-amino-9H-purin-9-yl)-3-fluoro-5,10,13-trihydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecin-9-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared on solid phase according to the methoddescribed above without capping steps.

ESI-MS (M-H)⁻ for C₂₁H₂₄FN₁₀O₁₂P₂ calculated: 689.1; found: 689.1; HPLCMethod 2, retention time (min)=40.4. ¹H and ³¹P NMR data in D₂O at 25°C.

H-1' H-2' H-3' H-4' H-5'a H-5'b P-CH₂-O H-2 H-8 ³¹P A 6.43 d 5.53 dd4.91 m 4.55 m 4.52 ddd 4.31 dt — 8.22 s 8.40 −1.74 (16.0) (51.7; (23.8;(9.0; 2.0; (12.2; (12.2; s 3.8) 9.0; 3.8) 1.6) 4.8; 2.0) 1.6; 1.6) G5.88 d 4.69 dd 4.48 dd 4.41 m 4.33 dt 4.165 bd 4.035 dd — 8.02 20.26(3.0) (4.9; (7.0; 4.9) (7.0; 2.2; (11.5; (11.5; 3.78 dd s 3.0) 1.8) 2.2;2.2) 3.5; 1.8) (15.0; 5.2) (15.0; 7.5)

Example 5.2-amino-9-((2R,3R,3aR,7aR,9R,10R,10aS,15aR)-9-(6-amino-9H-purin-9-yl)-3-fluoro-5,10,13-trihydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecin-2-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared on solid phase according to the methoddescribed above without capping steps.

ESI-MS (M-H)⁻ for C₂₁H₂₄FN₁₀O₁₂P₂ calculated: 689.1; found: 689.1; HPLCMethod 2, retention time (min)=37.8. ¹H and ³¹P NMR data in D₂O at 25°C. (coupling constants in parentheses where appropriae).

H-1' H-2' H-3' H-4' H-5'a H-5'b P-CH₂-O H-2 H-8 ³¹P G 6.21 d 5.47 dd4.85 4.50 md 4.48 m 4.31 br dt — — 7.95 −1.89 (15.8) (51.4; (overlap,(12.0; 1.5; s 3.8) HDO) 1.5) A 6.08 d 4.73 dd 4.46 m 4.49 m 4.39 br 4.21ddd 4.02 dd 8.19 s 8.40 20.16 (1.9) (4.4; 1.8) dt (11.7; 3.6; 3.75 dd s(11.7; 1.4) (15.1; 1.8; 1.8) 5.6) (15.1; 7.0)

Example 6.9,9′-((2R,3R,3aS,8aR,10R,11R,11aS,16aR)-3,6,11,14-tetrahydroxy-6,14-dioxidododecahydrodifuro[3,2-e:3′,2′-l][1,4,8,11]tetraoxa[2,9]diphosphacyclotetradecine-2,10-diyl)bis(2-amino-1,9-dihydro-6H-purin-6-one)

The product was prepared on solid phase according to the methoddescribed above without capping steps.

ESI-MS (M-H)⁻ for C₂₂H₂₇N₁₀O₁₄P₂ calculated: 717.1; found: 717.1; HPLCMethod 1, retention time (min)=31.5.

Example 7.2-amino-9-((2R,3R,3aS,7aR,9R,10R,10aR,15aR)-9-(6-amino-9H-purin-9-yl)-10-fluoro-3,5,13-trihydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecin-2-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared on solid phase according to the methoddescribed above without capping steps.

ESI-MS (M-H)⁻ for C₂₁H₂₄FN₁₀O₁₂P₂ calculated: 689.1; found: 689.1; HPLCMethod 1, retention time (min)=44.1. ¹H, ¹⁹F and ³¹P NMR data in D₂O at25° C.

Res. H-1' H-2' H-3' H-4' H-5'a H-5'b H-2 H-8 ¹⁹F ³¹P G 5.915 4.64 dd4.71 m 4.47 um 4.46 uM 4.24 ddd — 7.99 −199.16 −1.08 d (4.6; 2.3) (7.4;4.6) (12.0; s (2.3) 3.0; 1.2) A 6.37 d 5.58 dd 4.645 ddd 4.48 dt 4.41ddd 4.335 dt 8.19 s 8.36 — 19.73 (16.2) (51.8; (24.2; 9.0; (9.0; 2.6;(11.8; (11.8; s 3.6) 3.6) 2.4) 4.2; 2.6) 2.4; 2.4)

Example 8.2-amino-9-((2R,3R,3aR,7aR,9R,10R,10aR,15aR)-9-(6-amino-9H-purin-9-yl)-3,10-difluoro-5,13-dihydrox-5,13-diox-idodecahdro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecin-2-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared on solid phase using MOS oligonucleotidesynthesizer according to the method described above in 1 μmol scale onone column without capping steps.

CE-CPG was treated with 3% DCA in DCM for 2 min and then washed with ACNand DCM. A mixture of phosphonate Intermediate 6 in pyridine (0.1M, 0.1mL) and TIPSCl in pyridine (0.3M, 0.1 mL) was added, the mixture wasleft to react for 10 min and then washed with ACN and DCM.

The CPG was treated with 3% DCA in DCM for 2 min and then washed withACN and DCM. A mixture of phosphoramidite monomer 5 in acetonitrile(0.1M, 0.1 mL) and ETT in acetonitrile (0.3M, 0.1 mL) was added, themixture was left to react for 10 min and then washed with ACN and DCM.The CPG was then treated with a mixture of 5.5M tBuOOH in decane and DCM(1/4, v/v) for 3 min and subsequently washed with ACN and DCM. The CPGwas subsequently treated with 3% DCA in DCM for 2 min and then washedwith ACN and DCM.

A column containing the CPG-bound linear dimer was removed from thesynthesizer and CPG was transferred into 2 mL screw tube. 10% Solutionof Et₂NH in ACN (500 μl) was added to the CPG solid support with boundlinear dimer, and the heterogeneous mixture was shaken on an EppendorfThermomixer comfort shaker for 2 h at r.t. The solvents were removed. Asolution of DMOCP (10 mg) in pyridine (400 μl) was added into the tube.The heterogeneous mixture was shaken for 2 h at r.t. Then, 10 μl of Et₃Nand 266 μl of water were added to the reaction mixture and the shakingwas continued for 16 hours at 65° C. The reaction mixture wasconcentrated.

50% Aqueous MeOH (600 μl) was added to the reaction mixture and thesolid support was separated from the solution by decantation and washedwith 50% aqueous MeOH (3×200 μl). Solutions were collected andevaporated in 2 mL screw tube.

The residue was dissolved in 50% aqueous MeOH (600 μl) and a solution of33% MeNH₂ in EtOH (600 μl) was added. The mixture was shaken for 8 hoursat r.t. The solvents were then removed. The residue was dissolved in0.1M TEAB (1.5 ml) and applied to a semipreparative C18 column (Luna 5μm C18 250×10 mm). The desired product was isolated by reverse phasechromatography using linear gradient of acetonitrile in 0.1M TEAB:ESI-MS (M-H)⁻ for C₂₁H₂₃F₂N₁₀O₁₁P₂ calculated: 691.1; found: 691.1; HPLCMethod 2, retention time (min)=42.4. ¹H, ¹⁹F and ³¹P NMR data in D₂O at25° C. (coupling constants in parentheses where available).

Res. H-1' H-2' H-3' H-4' H-5'a H-5'b H-2 H-8 ¹⁹F ³¹P G 6.21 d 5.48 dd4.87 m 4.50 m 4.455 m 4.21 dd — 7.945 −199.00 −1.44 (15.6) (51.4; (11.6;s 3.8) 2.8) A 6.365 5.59 dd 4.62 ddd 4.48 m 4.48 m 4.285 br 8.16 s 8.36s −199.44 19.63 d (51.8; (24.0; 9.0; d (16.0) 3.6) 3.6) (11.8; <2)

Example 9.(2R,3S,3aS,8aR,10R,11S,11aS,16aR)-2,10-bis(6-amino-9H-purin-9-yl)-3,6,11,14-tetrahydroxvdodecahydrodifuro[3,2-e:3′,2′-l][1,4,8,11]tetraoxa[2,9]diphosphacyclotetradecine6,14-dioxide

The product was prepared on solid-phase according to the methoddescribed above without capping steps.

ESI-MS (M-H)⁻ for C₂₂H₂₇N₁₀O₁₂P₂ calculated: 685.1; found: 685.1; HPLCMethod 1, retention time (min)=30.6.

Example 10.(2R,3R,3aR,8aR,10R,11R,11aR,16aR)-2,10-bis(6-amino-9H-purin-9-yl)-3,6,11,14-tetrahydroxvdodecahydrodifuro[3,2-e:3′,2′-l][1,4,8,11]tetraoxa[2,9]diphosphacyclotetradecine6,14-dioxide

The product was prepared according to the method described above.

ESI-MS (M-H)⁻ for C₂₂H₂₇N₁₀O₁₂P₂ calculated: 685.1; found: 685.1; HPLCMethod 1, retention time (min)=30.8.

Example 11.2-amino-9-((2R,3R,3aS,7aR,9R,10R,10aR,15aR)-2-(6-amino-9H-purin-9-yl)-10-fluoro-3,5,13-trihydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10][tetraoxa][2,8]diphosphacyclotridecin-9-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared on solid-phase according to the methoddescribed above without capping steps

ESI-MS (M-H)⁻ for C₂₁H₂₄FN₁₀O₁₂P₂ calculated: 689.1; found: 689.1; HPLCMethod 2, retention time (min)=41.6. ¹H, ¹⁹F and ³¹P NMR data in D₂O at25° C.: ¹H NMR δ 8.42 (s, 1H), 8.21 (s, 1H), 7.80 (s, 1H), 6.18 (d,J=17.3 Hz, 1H), 6.13 (d, J=2.2 Hz, 1H), 5.51 (dd, J=52.1, 3.9 Hz, 1H),4.75 (m, 1H), 4.69 (dd, J=4.7, 2.2 Hz, 1H), 4.68 (ddd, J=21.0, 8.6, 3.9Hz, 1H), 4.51 (m, 1H), 4.44 (ddd, J=12.0, 4.3, 2.5 Hz, 1H), 4.42 (m,1H), 4.40 (dt, J=11.6, 2.4 Hz, 1H), 4.37 (dt, J=12.0, 2.4 Hz, 1H), 4.22(ddd, J=11.6, 3.4, 1.6 Hz, 1H), 4.08 (dd, J=14.8, 6.3 Hz, 1H), 3.91 (dd,J=14.8, 7.6 Hz, 1H); ¹⁹F NMR δ−198.94; ³¹P NMR δ 19.15, −1.37.

Example 12.2-amino-9-((2R,3R,3aR,7aR,9R,10R,10aR,15aR)-2-(6-amino-9H-purin-9-yl)-3,10-difluoro-5,13-dihydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecin-9-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared on solid phase using the MOS oligonucleotidesynthesizer according to the method described above on one column in 1μmol scale without capping steps.

CE-CPG was treated with 3% DCA in DCM for 2 min and then washed with ACNand DCM. A mixture of phosphonate Intermediate 7 in pyridine (0.1M, 0.1mL) and TIPSCl in pyridine (0.3M, 0.1 mL) was added, the mixture wasleft to react for 10 min and then washed with ACN and DCM.

The CPG was treated with 3% DCA in DCM for 2 min and then washed withACN and DCM. A mixture of phosphoramidite Intermediate 4 in acetonitrile(0.1M, 0.1 mL) and ETT in acetonitrile (0.3M, 0.1 mL) was added, and theresulting mixture was aged for 10 min and then washed with ACN and DCM.The CPG was treated with a mixture of 5.5M tBuOOH in decane and DCM(1/4, v/v) for 3 min and then washed with ACN and DCM. The CPG wastreated with 3% DCA in DCM for 2 min and then washed with ACN and DCM.

A column containing the CPG-bound linear dimer was removed from thesynthesizer and CPG was transferred into 2 mL screw tube. 10% Solutionof Et₂NH in ACN (500 μl) was added to the CPG solid support with boundlinear dimer, and the heterogeneous mixture was shaken on an EppendorfThermomixer comfort shaker for 2 h at r.t. The solvents were removed. Asolution of DMOCP (10 mg) in pyridine (400 μl) was then added into thetube. The heterogeneous mixture was shaken for 2 h at r.t. Then, 10 μlof Et₃N and 266 μl of water were added to the reaction mixture and theshaking was continued for 16 hours at 65° C. The reaction mixture wasconcentrated on Speed Vac.

50% Aqueous MeOH (600 μl) was added to the reaction concentrate and thesolid support was separated from the solution by decantation and washedwith 50% aqueous MeOH (3×200 μl). Solutions were collected andconcentrated in 2 mL screw tube. The residue was dissolved in 50%aqueous MeOH (600 μl) and a solution of 33% MeNH₂ in EtOH (600 μl) wasadded. The mixture was shaken for 8 hours at r.t. before the solventswere removed. The residue was dissolved in 0.1M TEAB (1.5 ml) andpurified on a semipreparative C18 column (Luna 5 μm C18 250×10 mm). Thedesired product was isolated by reverse phase chromatography usinglinear gradient of acetonitrile in 0.1M TEAB: ESI-MS (M-H)⁻ forC₂₁H₂₃F₂N₁₀O₁₁P₂ calculated: 691.1; found: 691.1; HPLC Method 2,retention time (min)=43.2. ¹H, ¹⁹F and ³¹P NMR data in D₂O at 25° C.: ¹HNMR δ 8.38 (s, 1H), 8.18 (s, 1H), 7.97 (s, 1H), 6.43 (d, J=15.6 Hz, 1H),6.19 (d, J=17.0 Hz, 1H), 5.53 (dd, J=51.7, 4.0 Hz, 1H), 5.51 (dd,J=51.4, 4.0 Hz, 1H), 4.89 (ddd, J=21.0, 9.0, 4.0 Hz, 1H), 4.63 (ddd,J=22.5, 8.8, 4.0 Hz, 1H), 4.55 (m, 1H), 4.52 (ddd, J=12.1, 4.4, 2.1 Hz,1H), 4.42 (m, 1H), 4.39 (dt, J=11.8, 2.4 Hz, 1H), 4.33 (dt, J=12.1, 1.8Hz, 1H), 4.19 (ddd, J=11.8, 3.8, 1.8 Hz, 1H), 4.08 (dd, J=14.6, 5.5 Hz,1H), 3.86 (dd, J=14.6, 8.8 Hz, 1H); ¹⁹F NMR δ −198.84, −199.40; ³¹P NMRδ 19.05, −1.80.

Example 13.(2R,3R,3aR,7aR,9R,10R,10aR,15aR)-2,9-bis(6-amino-9H-purin-9-yl)-3,10-difluoro-5,13-dihydroxydecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecine5,13-dioxide

The product was prepared on solid-phase according to the methoddescribed above without capping steps.

Alternatively, the desired product was prepared in solution according tothe following protocol:

ETT (0.1 g; 0.82 mmol) was added to a solution of phosphonateIntermediate 9 (0.1 g; 0.17 mmol) and phosphoramidite Intermediate 10(cat. #203-51 purchased from Metkinen, 0.16 g; 0.19 mmol) in DCM (10mL). The reaction mixture was stirred under argon for 2 h at r.t. Afterthat, CSO (0.15 g; 0.56 mmol) was added and the mixture was stirred for1 h at r.t. After that, the reaction mixture was treated with methanol(10 mL) for 1 h at r.t. and evaporated. The residue was treated with 80%acetic acid in water (10 mL) for 2 h at r.t. The solution was directlyloaded on C18 column and the linear phosphonate dimer Intermediate 11was isolated using linear gradient of acetonitrile in 5 mM ammoniumbicarbonate. Intermediate 11: ESI-MS calcd for C₄₅H₄₃F₂N₁₂O₁₄P₂ (M-H)⁻1075.3, found 1075.3.

DMOCP (0.16 g; 0.85 mmol) was added a solution of linear phosphonatedimer Intermediate 11 in pyridine (6 mL) and the reaction mixture wasstirred for 4 h at r.t. After that, water (4 mL) was added and themixture was stirred for 3 h at 65° C., evaporated and coevaporated withmethanol (3×10 mL). The residue was treated with 10% DEA/ACN for 2 h atr.t. and evaporated. Cyclic dinucleotide was isolated using lineargradient of acetonitrile in 5 mM ammonium bicarbonate and evaporated.The residue was dissolved in 50% ACN in water (10 mL) and 33%methylamine in ethanol (5 mL) was added. The mixture was stirred for 2 hand evaporated. The product was purified using preparative C18 HPLC(elution with linerar gradient of acetonitrile in 0.1M TEAB). Thedesired product was converted to sodium salt using DOWEX Na⁺ andfreeze-dried from water: ESI-MS (M-H)⁻ for C₂₁H₂₃F₂N₁₀O₁₀P₂ calculated:675.1; found: 675.1; HPLC Method 2, retention time (min)=43.1. ¹H, ¹⁹Fand ³¹P NMR data in D₂O at 25° C.: ¹H NMR δ 8.36 (s, 1H), 8.31 (s, 1H),8.20 (s, 1H), 8.19 (s, 1H), 6.41 (d, J=15.9 Hz, 1H), 6.38 (d, J=15.3 Hz,1H), 5.54 (dd, J=51.4, 3.8 Hz, 1H), 5.44 (dd, J=51.8, 3.7 Hz, 1H), 4.86(m, 1H), 4.56 (m, 1H), 4.56 (ddd, J=9.0, 2.2, 1.6 Hz, 1H), 4.53 (m, 1H),4.49 (m, 1H), 4.47 (dt, J=12.0, 2.2 Hz, 1H), 4.33 (dt, J=11.8, 1.6 Hz,1H), 4.24 (ddd, J=12.0, 3.4, 1.6 Hz, 1H), 4.09 (dd, J=14.9, 5.4 Hz, 1H),3.84 (dd, J=14.9, 8.4 Hz, 1H); ¹⁹F NMR δ −198.64, −199.19; ³¹P NMR δ19.46, −2.05.

Example 14.2-amino-9-((2R,3R,3aR,7aR,9R,10aS,15aR)-2-(6-amino-9H-purin-9-yl)-3-fluoro-5,13-dihydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecin-9-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared on solid-phase according to the methoddescribed above without capping steps.

ESI-MS (M-H)⁻ for C₂₁H₂₄FN₁₀O₁₁P₂ calculated: 673.1; found: 673.1; HPLCMethod 2, retention time (min)=39.5.

Example 15.2-amino-9-((2R,3aS,7aR,9R,10R,10aR,15aR)-9-(6-amino-9H-purin-9-yl)-10-fluoro-5,13-dihydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecin-2-yl)-1,9-dihydro-6H-purin-6-one

The product was prepared on solid-phase according to the methoddescribed above without capping steps.

ESI-MS (M-H)⁻ for C₂₁H₂₄FN₁₀O₁₁P₂ calculated: 673.1; found: 673.1; HPLCMethod 2, retention time (min)=47.5.

Example 16.9,9′-((2R,3R,3aR,7aR,9R,10R,10aR,15aR)-3,10-difluoro-5,13-dihydroxy-5,13-dioxidodecahydro-2H-difuro[3,2-d:3′,2′-k][1,3,7,10]tetraoxa[2,8]diphosphacyclotridecine-2,9-diyl)bis(2-amino-1,9-dihydro-6H-purin-6-one)

The product was prepared on solid-phase according to the methoddescribed above without capping steps.

ESI-MS (M-H)⁻ for C₂₁H₂₃F₂N₁₀O₁₂P₂ calculated: 707.1; found: 707.1; HPLCMethod 2, retention time (min)=37.1.

Example 17. Biological Evaluation

A cyclic dinucleotide was determined to be a STING agonist: (A) if itdemonstrated binding to the AQ allelic form of human STING protein withthermal shift of >0.5° C. in the STING Differential Scanning FluorimetryAssay (DSF), and (B) if it demonstrated STING activation through IRF-3dependent expression of firefly luciferase reporter with EC₅₀<100μmol·l⁻¹.

ISRE Reporter Plasmid (pGL64.27-4×ISRE)

Two complementary oligonucleotides of the sequence AAAGATCTTGGAAAGTGAAACCTTGGAAAAC GAAACTGGACAAAGGGAAACTG CAGAAACTGAAACAAAGCTTAA (SEQ ID NO: 1)and TTAAGCTTTGTTTCAGTTTCTGCAGTTTCCCTTTGTCCAGTTTCGTTTTCCAAGGTTTCACTTTCCAAGATCTTT (SEQ ID NO:2) containing four interferon-sensitiveresponse elements (ISRE) were synthesized by Sigma Aldrich (CzechRepublic, Prague). The oligonucleotides were mixed in equal molaramounts, hybridized, and cleaved by restriction endonucleases HindIII(cat. #. R0104S, NEB, Ipswich, USA) and BglII (cat. #R0144S, NEB,Ipswich, USA). Ultimately, they were ligated into plasmid pGL4.27 (cat.#E6651, Promega, Madison, USA) linearized with the same enzymes. Asresult the sequence with four ISRE sites was placed upstream of theminimum promoter of firefly luciferase reporter gene.

293T wtSTING-FL Reporter Cells

293T cells (cat. #CRL-3216, ATCC, Manassas, USA) were seeded a daybefore transfection at density 125,000 cells per cm² onto poly-D-lysine(cat. #P6407, Sigma Aldrich, Czech Republic) coated six well plates inantibiotic free DMEM with high glucose (cat. #D5796, Sigma Aldrich,Czech Republic) supplemented with 10% heat inactivated FBS (cat. #S1520,Biowest, Riverside, USA). On the day of transfection, 2.5 μg of theplasmid pUNO1-hSTING-WT (cat. #punol-hstingwt, InvivoGen, San Diego,USA,) encoding human wild type STING (WT STING) was diluted in 125 μLOptiMEM medium (cat. #31985062, ThermoFisher, Waltham, USA) and mixedwith 125 μL of the same medium containing 12.5 μL of Lipofectamine 2000(cat. #11668019, ThermoFisher, Waltham, USA). After 5 minutes incubationat room temperature (RT), 250 μL of the mixture was added dropwise tothe cells in one well. Cells were incubated 36 hours at 37° C. with 5%CO₂, and then detached with 0.05% Trypsin and 0.22 g/L EDTA (both cat.#L0941, Biowest, Riverside, USA).

Transfected cells were seeded onto poly-D-lysine coated six well platesat density 50,000 cells per 1 cm² in DMEM medium with high glucosecontaining 10% heat inactivated FBS, 30 μg/mL blasticidin (cat.#ant-bl-05, InvivoGen, San Diego, USA), 0.06 mg/ml Penicillin G and 0.1mg/ml Streptomycin Sulfate (both cat. #. L0018, Biowest, Riverside,USA). The medium was replenished every 3-4 days until visible coloniesof cells resistant to blasticidin were formed.

Blasticidin resistant cells stably expressing WT STING were furthertransfected with pGL64.27-4×ISRE plasmid following the same procedure asdescribed above. The transfected cells were selected for the resistanceto 300 μg/mL hygromycin (cat. #. 10687010, ThermoFisher, Waltham, USA)in DMEM with high glucose containing 10% heat inactivated FBS, 30 μg/mLblasticidin, 0.06 mg/ml Penicillin G and 0.1 mg/ml Streptomycin Sulfate.Homogeneous culture of stably double transfected cells was prepared bylimiting dilution of cells in 96 well plates and wells with cells wereselected that originated from a single cell. These cells were expanded,and expression of WT STING was confirmed by western blot usingmonoclonal mouse anti-STING antibodies (cat. #. MAB7169, 1:1000dillution; 2° antibody cat. #. HAF007, 1:2000 dilution, both from R&DSystems, Minneapolis, USA), and by induction of firefly luciferaseexpression in the presence of 50 μM STING agonist 2′3′ cGAMP (cat.#tlrl-nacga23, InvivoGen, San Diego, USA). Genomic DNA from thetransfected cells was amplified with primers pUNO1_Seq_F(TGCTTGCTCAACTCTACGTC) (SEQ ID NO:3) and pUNO1_Seq_R(GTGGTTTGTCCAAACTCATC) (SEQ ID NO:4) that were complementary to pUNO1plasmid and the presence of WT STING gene in the transfected cells wasconfirmed by DNA sequencing.

Digitonin Assay Using 293T wtSTING-FL Reporter Cells

293T wtSTING-FL cells were seeded at density of 250,000 cells per cm²onto 96 well poly-D-lysine coated plates in 100 μl DMEM with highglucose supplemented with 10% heat inactivated FBS. The medium wasremoved next day and three fold serial dilutions of compounds inDigitonin buffer containing 50 mmol·l⁻¹ HEPES (cat. #H3375, SigmaAldrich, Czech Republic) pH 7.0, 100 mmol·l⁻¹ KCl, 3 mmol·l⁻¹ MgCl₂, 0.1mmol·l⁻¹ DTT (cat. #D0632, Sigma Aldrich, Czech Republic), 85 mmol·l⁻¹Sucrose (cat. #S7903, Sigma Aldrich, Česká Republika), 0.2% BSA (cat.#A2153, Sigma Aldrich, Czech Republic), 1 mmol·l⁻¹ ATP (cat. #A1852,Sigma Aldrich, Czech Republic), 0.1 mmol·l⁻¹ GTP (cat. #G8877, SigmaAldrich, Czech Republic), and 10 μg/mL Digitonin A (cat. #D141, SigmaAldrich, Czech Republic) were added to the cells. The buffer was removedafter 30 minutes incubation at 37° C. with 5% CO₂, the cells were washedonce with 100 μl of cultivation medium, and 100 μl of medium was addedto each well. The plates with cells were incubated for 5 hours at 37° C.with 5% CO₂, 50 μl of the medium was removed and 30 μl of ONE-Glo™Luciferase Assay System reagent (cat. #E6120, Promega, Madison, USA) wasadded to each well. Luminesce was read on Synergy H1 (Biotek, Winooski,USA). GraphPad Prism (San Diego, Calif., USA) was used to calculate the50% effective concentration (EC₅₀) from an 8-point dose-response curve.Control compounds 3′3′-c-di-GMP (cat. #tlrl-nacdg), 3′3′-c-di-AMP (cat.#tlrl-nacda), 3′3′-cGAMP (cat. #tlrl-nacga), 2′3′-cGAMP (cat.#tlrl-nacga23), and 2′2′-cGAMP (cat. #tlrl-nacga22) were purchased fromInvivogen (San Diego, USA).

WT STING and AQ STING Proteins

Both WT and AQ human STING (G230A-R293Q) cDNA were amplified by the useof PCR (Phusion® High-Fidelity DNA Polymerase, cat. #M0530S, NEB,Ipswich, USA) using oligonucleotides hSTING140-BamH-For(GTGGGATCCGCCCCAGCTGAGATCTCTGCAG) (SEQ ID NO:5) and hSTING379-Not-Rev3(TATGCGGCCGCCTATTACACAGTAACCTCTTCCTTTTC) (SEQ ID NO:6) frompUNO1-hSTING-WT (cat. #punol-hstingwt, InvivoGen, San Diego, USA) andpUNO1-hSTING-HAQ plasmids (punol-hsting-haq, InvivoGen, San Diego, USA).Purified PCR products were cleaved with restriction enzymes BamHI (cat.#R0136S, NEB, Ipswich, USA) and NotI (cat. #R0189S, NEB, Ipswich, USA)and cloned into the pSUMO vector linearized with the identical enzymes.Plasmid pSUMO was created by introducing 8-His-SUMO sequence betweenNdeI and BamHI sites of pHis-parallel2 plasmid (Clontech, Moutain View,USA). pSUMO-STING WT or pSUMO-STING AQ plasmids thus encoded truncatedhuman WT STING or AQ STING (amino acid residues 140-343) with N-terminal8×His and SUMO tag.

The recombinant WT STING and AQ STING proteins were overexpressed inRosetta-gami B (DE3) competent cells (cat. #71136-3, Merck Millipore,Billerica, USA). Bacterial pellets were re-suspended in ice-cold lysisbuffer containing 50 mmol·l⁻¹ TrisCl (cat. #T1503, Sigma Aldrich, CzechRepublic) pH 8.0, 300 mmol·l⁻¹ NaCl, 3 mmol·l⁻¹ β-mercaptoethanol (cat.#M6250, Sigma Aldrich, Czech Republic), 10% glycerol (cat. #G5516, SigmaAldrich, Czech Republic) and 20 mmol·l⁻¹ imidazole (cat. #15513, SigmaAldrich, Czech Republic) using Dounce homogenizer. DNase I (cat. #D5025,Sigma Aldrich, Czech Republic) and RNase A (cat. #R6513, Sigma Aldrich,Czech Republic) were added (final concentration 50 μg/ml) together withMgCl₂ (final concentration 5 mmol·l⁻¹) to the homogenate and bacteriawere lysed using French Press G-M™ High-Pressure Cell Press Homogenizer(1500 psi, 3 cycles). Lysate was spun 30,000 g for 20 minutes andsupernatant was gently stirred with Ni-NTA resin (cat. #745400.25Macherey-Nagel, Düren, Germany) for 30 minutes. The resin was pouredinto a chromatography column, washed with 50 ml buffer A (50 mmol·l⁻¹TrisCl (pH 8.0), 800 mmol·l⁻¹ NaCl, 3 mmol·l⁻¹ β-mercaptoethanol; 10%glycerol; 20 mmol·l⁻¹ imidazole) and 8-His-SUMO tagged STING proteinswere eluted with 15 ml buffer A containing 300 mmol·l⁻¹ imidazole. Theeluted proteins were cleaved with recombinant SUMO protease (80 μg/ml ofprotein solution, cat. #12588018, ThermoFisher, Waltham, USA). Theproteins were further purified by size exclusion chromatography usingHiLoad 16/60 Superdex 75 (cat. #28989333, GE Healthcare Bio-Sciences,Pittsburgh, USA) in 50 mmol·l⁻¹ Tris Cl buffer pH 7.4 containing 150mmol·l⁻¹ NaCl, and 10% glycerol. Proteins were concentrated with Amicon®Ultra-15 10 K device (cat. #UFC901008, Merck Millipore, Billerica, USA)and flash frozen in liquid N₂.

DNA sequence of 8-His-SUMO (SEQ ID NO: 7)ATGTCGCATCACCATCATCATCACCACCATGGGATGTCGGACTCAGAAGTCAATCAAGAAGCTAAGCCAGAGGTCAAGCCAGAAGTCAAGCCTGAGACTCACATCAATTTAAAGGTGTCCGATGGATCTTCAGAGATCTTCTTCAAGATCAAAAAGACCACTCCTTTAAGAAGGCTGATGGAAGCGTTCGCTAAAAGACAGGGTAAGGAAATGGACTCCTTAAGATTCTTGTACGACGGTATTAGAATTCAAGCTGATCAGACCCCTGAAGATTTGGACATGGAGGATAACGATATTATTGAGGCTCACCGCGAACAGATTGGTGGATCC.Amino acid sequence of 8-His-SUMO (SEQ ID NO: 8)MSHHHHHHHHGMSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMED NDIIEAHREQIGGS.Amino acid sequence of truncated WT STING (SEQ ID NO: 9)APAEISAVCEKGNFNVAHGLAWSYYIGYLRLILPELQARIRTYNQHYNNLLRGAVSQRLYILLPLDCGVPDNLSMADPNIRFLDKLPQQTGDRAGIKDRVYSNSIYELLENGQRAGTCVLEYATPLQTLFAMSQYSQAGFSREDRLEQAKLFCRTLEDILADAPESQNNCRLIAYQEPADDSSFSLSQEVLR HLRQEEKEEVTV.Amino acid sequence of truncated AQ STING (SEQ ID NO: 10)APAEISAVCEKGNFNVAHGLAWSYYIGYLRLILPELQARIRTYNQHYNNLLRGAVSQRLYILLPLDCGVPDNLSMADPNIRFLDKLPQQTADRAGIKDRVYSNSIYELLENGQRAGTCVLEYATPLQTLFAMSQYSQAGFSREDRLEQAKLFCQTLEDILADAPESQNNCRLIAYQEPADDSSFSLSQEVLR HLRQEEKEEVTV.Differential Scanning Fluorimetry with WT STING and AQ STING

WT and AQ allelic forms of STING protein were diluted to the finalconcentration 0.1 mg/ml in 100 mmol·l⁻¹ TrisCl buffer pH 7.4 containing,150 mmol·l⁻¹ NaCl, 1:500 SYPRO Orange (cat. #S6650, ThermoFisher,Waltham, USA) and 150 μM CDN or water. 20 μL solutions of the reactionmixtures were pipetted in triplicates into 96 well optical reactionplates and thermal denaturation of samples were performed on real timePCR cycler (LightCycler® 480 Instrument II—Roche, Basel, Switzerland).The first derivative of the thermal denaturation curves was performed tocalculate denaturing temperatures of STING-CDN complexes and STINGapoproteins. The thermal shift for each CDN was calculated bysubtracting the average denaturing temperature of STING apoprotein fromthe average denaturing temperature of STING CDN complex.

TABLE 2 Biological evaluation of CDNs Digitonin assay DSF ΔTm (° C.)EC₅₀ (μmol · l⁻¹) Compound WT STING AQ STING WT STING Example 2 0.1 3.310.4 Example 3 0.0 3.1 12.5 Example 4 4.8 13.6 0.05 Example 5 4.1 12.00.1 Example 6 0.1 0.0 >50 Example 7 5.6 15.0 0.03 Example 8 10.7 18.60.008 Example 9 0.1 0.1 >100 Example 10 0.0 −0.2 >100 Example 11 7.311.3 0.05 Example 12 9.3 17.3 0.02 Example 13 9.4 16.1 0.012 Example 143.8 11.2 0.09 Example 15 4.0 12.7 0.11 Example 16 3.1 10.7 0.33′3′-c-di-GMP 2.6 7.7 5.8 3′3′-c-di-AMP 2.6 9.3 0.3 3′3′-cGAMP 5.1 13.30.6 2′2′-cGAMP 11.5 19.4 0.03 2′3′-cGAMP 15.2 22.7 0.03

Although the foregoing invention has been described in some detail byway of illustration and Example for purposes of clarity ofunderstanding, one of skill in the art will appreciate that certainchanges and modifications may be practiced within the scope of theappended claims. In addition, each reference provided herein isincorporated by reference in its entirety to the same extent as if eachreference was individually incorporated by reference. Where a conflictexists between the instant application and a reference provided herein,the instant application shall dominate.

What is claimed is:
 1. A compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein L¹ is —O— or—K—C(R⁶R⁷)—; L² is —K—C(R⁶R⁷)—; Y¹ and Y² are each independently —O—,—S—, or —CH₂—; X¹ and X³ are each independently OH, SH, OR¹³, SR¹³, orN(R¹³)₂; X² and X⁴ are each independently O or S; R¹, R⁵, R⁸ and R¹² areeach independently H, CH₂F, CHF₂, CF₃, CN, N₃, F, Cl, Br, I, COOR¹³,CON(R¹³)₂, CH₂OH, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆substituted alkyl, C₂-C₆ substituted alkenyl, C₂-C₆ substituted alkynyl,OR¹³, SR¹³, or N(R¹³)₂; R², R³, R⁴, R⁹, R¹⁰ and R¹¹ are eachindependently H, OH, F, Cl, Br, I, CN, N₃, C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₁-C₆ substituted alkyl, C₂-C₆ substituted alkenyl, C₂-C₆substituted alkynyl, OR¹³, SR¹³, or N(R¹³)₂; R⁶ and R⁷ are eachindependently H, CH₂F, CHF₂, CF₃, CN, N₃, F, Cl, Br, I, COOR¹³,CON(R¹³)₂, CH₂OH, CH₂N₃, OR¹³, SR¹³, N(R¹³)₂, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₁-C₆ substituted alkyl, C₂-C₆ substitutedalkenyl, C₂-C₆ substituted alkynyl, C₃-C₇ cycloalkyl, C₂-C₁₀heterocycloalkyl, C₂-C₁₀ substituted heterocycloalkyl, C₆-C₁₀ aryl,C₆-C₁₀ substituted aryl, C₂-C₁₀ heteroaryl, or C₂-C₁₀ substitutedheteroaryl; each R¹³ is independently H, —C(═Z)R¹⁴, —C(═Z)OR¹⁴,—C(═Z)SR¹⁴, —C(═Z)N(R¹⁴)₂, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₁-C₆ substituted alkyl, C₂-C₆ substituted alkenyl, C₂-C₆ substitutedalkynyl, C₃-C₇ cycloalkyl, C₂-C₁₀ heterocycloalkyl, C₂-C₁₀ substitutedheterocycloalkyl, C₆-C₁₀ aryl, C₆-C₁₀ substituted aryl, C₂-C₁₀heteroaryl, or C₂-C₁₀ substituted heteroaryl; each R¹⁴ is independentlyH, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ substituted alkyl,C₂-C₆ substituted alkenyl, C₂-C₆ substituted alkynyl, C₃-C₇ cycloalkyl,C₂-C₁₀ heterocycloalkyl, C₂-C₁₀ substituted heterocycloalkyl, C₆-C₁₀aryl, C₆-C₁₀ substituted aryl, C₂-C₁₀ heteroaryl, or C₂-C₁₀ substitutedheteroaryl; each Z is independently O, S, or NR¹³; K is a variable thatrepresents O, S, S(O), S(O)₂, NH, or NR¹³; Base¹ and Base² are eachindependently:

wherein A, A¹, A², A³ and A⁴ are each independently H, OH, SH, F, Cl,Br, I, NH₂, OR¹³, SR¹³, NHR¹³, N(R¹³)₂, or R¹⁴; and wherein the C₁-C₆substituted alkyl, C₂-C₆ substituted alkenyl, C₂-C₆ substituted alkynyl,C₂-C₁₀ substituted heterocycloalkyl, C₆-C₁₀ substituted aryl, or C₂-C₁₀substituted heteroaryl is substituted by 1, 2, 3, or 4 substituentsindependently selected from the group consisting of —OH, —SH, —NH₂, ═O,═NH, ═S, ≡N, halogen, —N₃, C₁₋₆ alkyl, C₁₋₆ alkoxy, —CN and —COOR_(p),where R_(p) is hydrogen or C₁ to C₆ alkyl.
 2. The compound of claim 1,wherein Y¹ and Y² are O.
 3. The compound of claim 1, wherein L¹ is —O—or —O—C(R⁶R⁷)—, and L² is —O—C(R⁶R⁷)—.
 4. The compound of claim 1,wherein the compound of Formula (I) has a structure of Formula (Ia-2):

or a pharmaceutically acceptable salt thereof.
 5. The compound of claim4, wherein L¹ is —O— or —O—CH₂—.
 6. The compound of claim 1, wherein thecompound of Formula (I) has a structure of Formula (Ib):

or a pharmaceutically acceptable salt thereof.
 7. The compound of claim1, wherein the compound of Formula (I) has a structure of Formula (Ic):

or a pharmaceutically acceptable salt thereof.
 8. The compound of claim1, wherein the compound of Formula (I) has a structure of Formula (Id):

or a pharmaceutically acceptable salt thereof.
 9. The compound of claim1, wherein Base¹ and Base² are each independently:


10. The compound of claim 1, wherein Base¹ and Base² are eachindependently:


11. The compound of claim 1, wherein R³ and R¹⁰ are each independentlyH, OH, F, Cl, Br, I, CN, N₃, or OR¹³, wherein R¹³ is C₁-C₆ alkyl. 12.The compound of claim 1, wherein R³ and R¹⁰ are each independently H, OHor F.
 13. The compound of claim 1, wherein the compound has thestructure:

or a pharmaceutically acceptable salt thereof.
 14. A pharmaceuticalcomposition comprising the compound of claim 1, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier,excipient, and/or diluent.